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The list of standard abbreviations for JDS is available at adsa.org/jds-abbreviations-24. Nonstandard abbreviations are available in the Notes. During processing and storage of both conventional and lactose-hydrolyzed UHT milk (LHM), aggregation of milk proteins occurs. Protein aggregation can inter alia occur via nonreducible covalent cross-links derived from either Maillard or dehydroalanine (DHA) pathways. To study this further in relation to processing method and lactase enzyme purity, LHM was produced using 3 different lactase preparations, with lactase enzymes added in a dairy setting either before (prehydrolysis) or after (posthydrolysis) UHT treatment. The prepared LHM types were subsequently stored at either 25°C or 35°C for up to one year. Mass spectrometry was used to absolutely quantify the level of furosine, N-ε-(carboxymethyl)lysine, N-ε-(carboxyethyl)lysine, lanthionine, and lysinoalanine in these products using a newly developed method on triple quadrupole for these processing-induced markers. The results showed higher levels of Maillard-related processing markers in prehydrolyzed LHM compared with posthydrolyzed LHM and conventional UHT milk, which, on the other hand, contained higher concentrations of DHA-derived cross-links. Proteomics of collected particles from asymmetrical flow field-flow fractionation in combination with gel electrophoresis indicated the presence of intramicellar cross-links during storage, especially for larger particles involving α S1 - and α S2 -caseins. [ABSTRACT FROM AUTHOR]

Three main mechanisms of separation in Field-Flow Fractionation, the polarization, steric, and focusing, are critically analyzed. The theoretical evaluation of these mechanisms is accompanied by supporting data from several previous and recent experimental findings. The polarization mechanism can be exploited universally for the macromolecules in solution and for small particulate species. The steric mechanism is impracticable for the separations of the macromolecules in solution. The focusing mechanism was successfully applied in high-speed separations of particulate species, including the biological cells. The major factors differentiating three mechanisms and delimiting their practical use are the strength of the effective field force, the flow-rate of the carrier liquid, and the diffusion coefficients of the separated species. Whenever the highest selectivity is preferred, the polarization and focusing Field-Flow Fractionation, performed under optimal experimental conditions, are rivaling other separation methods. The accuracy, the reproducibility and the repeatability of the field-flow fractionation analyses was evaluated by simultaneous measurements carried out in two independent laboratories. [ABSTRACT FROM AUTHOR]

In recent years, the use of lipid nanoparticles (LNPs) for delivery of messenger RNA (mRNA)–based therapies has gained substantial attention in the field of drug development. In such an application, multiple LNP attributes have to be carefully characterized to ensure product safety and quality, whereas accurate and efficient characterization of these complex mRNA-LNP formulations remains a challenging endeavor. Here, we present the development and application of an online separation and characterization platform designed for the isolation and in-depth analysis of mRNAs and mRNA-loaded LNPs. Our asymmetrical flow field-flow fractionation with a multi-detector (MD-AF4) method has demonstrated exceptional resolution between mRNA-LNPs and mRNAs, delivering excellent recoveries (over 70%) for both analytes and exceptional repeatability. Notably, this platform allows for comprehensive and multi-attribute LNP characterization, including online particle sizing, morphology characterization, and determination of encapsulation efficiency, all within a single injection. Furthermore, real-time online sizing by synchronizing multi-angle light scattering (MALS) and dynamic light scattering (DLS) presented higher resolution over traditional batch-mode DLS, particularly in differentiating heterogeneous samples with a low abundance of large-sized particles. Additionally, our method proves to be a valuable tool for monitoring LNP stability under varying stress conditions. Our work introduces a robust and versatile analytical platform using MD-AF4 that not only efficiently provides multi-attribute characterizations of mRNA-LNPs but also holds promise in advancing studies related to formulation screening, quality control, and stability assessment in the evolving field of nanoparticle delivery systems for mRNAs. [ABSTRACT FROM AUTHOR]

The current spotlight of cancer therapeutics is shifting towards personalized medicine with the widespread use of monoclonal antibodies (mAbs). Despite their increasing potential, mAbs have an intrinsic limitation related to their inability to cross cell membranes and reach intracellular targets. Nanotechnology offers promising solutions to overcome this limitation, however, formulation challenges remain. These challenges are the limited loading capacity (often insufficient to achieve clinical dosing), the complex formulation methods, and the insufficient characterization of mAb-loaded nanocarriers. Here, we present a new nanocarrier consisting of hyaluronic acid-based nanoassemblies (HANAs) specifically designed to entrap mAbs with a high efficiency and an outstanding loading capacity (50%, w/w). HANAs composed by an mAb, modified HA and phosphatidylcholine (PC) resulted in sizes of ~ 100 nm and neutral surface charge. Computational modeling identified the principal factors governing the high affinity of mAbs with the amphiphilic HA and PC. HANAs composition and structural configuration were analyzed using the orthogonal techniques cryogenic transmission electron microscopy (cryo-TEM), asymmetrical flow field-flow fractionation (AF4), and small-angle X-ray scattering (SAXS). These techniques provided evidence of the formation of core-shell nanostructures comprising an aqueous core surrounded by a bilayer consisting of phospholipids and amphiphilic HA. In vitro experiments in cancer cell lines and macrophages confirmed HANAs' low toxicity and ability to transport mAbs to the intracellular space. The reproducibility of this assembling process at industrial-scale batch sizes and the long-term stability was assessed. In conclusion, these results underscore the suitability of HANAs technology to load and deliver biologicals, which holds promise for future clinical translation. [ABSTRACT FROM AUTHOR]

Due to very high mobility in the environment and penetration ability into living organisms, nanoparticles (NPs) of urban dust pose a potential threat to human health and urban ecosystems. Currently, data on the chemical composition of NPs of urban dust, their fate in the environment, and corresponding risks are rather limited. In the present work, NPs of deposited urban dust have been comprehensively studied for the first time; NPs isolated from 78 samples of dust collected in Moscow, the largest megacity in Europe, being taken as example. The elemental composition, potential sources as well as environmental, ecological, and health risks of NPs of urban dust are assessed. It is found that dust NPs are extremely enriched by Cu, Hg, Zn, Mo, Sb, and Pb, and can serve as their carrier in urban environments. No regularities in the spatial distribution of elements have been found, probably, due to high mobility of dust NPs. High ecological and health risks caused by dust NPs are demonstrated. Source apportionment study has evaluated one natural and two anthropogenic sources of elements in NPs of urban dust; the contribution of natural and anthropogenic sources being comparable. It is also shown that dust NPs may be considered as an important carrier of trace elements in urban aquatic systems. Additionally, the risks associated with NPs and bulk samples of dust have been compared. The observed risks associated with NPs are significantly higher. [ABSTRACT FROM AUTHOR]

ABSTRACT: During processing and storage of both conventional and lactose-hydrolyzed UHT milk (LHM), aggregation of milk proteins occurs. Protein aggregation can inter alia occur via nonreducible covalent cross-links derived from either Maillard or dehydroalanine (DHA) pathways. To study this further in relation to processing method and lactase enzyme purity, LHM was produced using 3 different lactase preparations, with lactase enzymes added in a dairy setting either before (prehydrolysis) or after (posthydrolysis) UHT treatment. The prepared LHM types were subsequently stored at either 25°C or 35°C for up to one year. Mass spectrometry was used to absolutely quantify the level of furosine, N-ε-(carboxymethyl)lysine, N-ε-(carboxyethyl)lysine, lanthionine, and lysinoalanine in these products using a newly developed method on triple quadrupole for these processing-induced markers. The results showed higher levels of Maillard-related processing markers in prehydrolyzed LHM compared with posthydrolyzed LHM and conventional UHT milk, which, on the other hand, contained higher concentrations of DHA-derived cross-links. Proteomics of collected particles from asymmetrical flow field-flow fractionation in combination with gel electrophoresis indicated the presence of intramicellar cross-links during storage, especially for larger particles involving αS1- and αS2-caseins.

The current absence of well-established and standardized methods for characterizing submicrometer- and nano-sized particles in water samples presents a significant analytical challenge. With the increasing utilization of nanomaterials, the potential for unintended exposure escalates. The widespread and persistent pollution of water by micro- and nanoplastics globally is a concern that demands attention, not only to reduce pollution but also to develop methods for analyzing these pollutants. Additionally, the analysis of naturally occurring nano entities such as bubbles and colloidal matter poses challenges due to the lack of systematic and consistent methodologies. This study presents Asymmetric Flow Field-Flow Fractionation (AF4) separation coupled with a UV-VIS spectrometer followed by Multi-Angle Light Scattering (MALS) for detection and size characterization of nanometric entities. It is coupled with an Inductively Coupled Plasma Mass Spectrometer (ICP-MS) for elemental analysis. Water samples from different sources, such as untreated mountain spring water, groundwater, and bottled drinking water, were analyzed. The system was calibrated using pure particle standards of different metallic compositions. Our study demonstrates the capability of AF4-UV-MALS-ICP-MS to detect metals such as Al, Ba, Cu, and Zn in particles of around 200 nm diameter and Mg associated with very small particles between 1.5 and 10 nm in different drinking water samples. [ABSTRACT FROM AUTHOR]

Synthetic amorphous silica (SAS) is used as additive in a variety of industrial applications for many decades and has been approved to be used in food, food contact materials, pharmaceuticals, and cosmetics. Due its internal structure, SAS is considered as a nanomaterial, thus it is affected by a general safety discussion. Based on the production process, SAS for cosmetic application is a nanomaterial by the EU Recommendation, although it was not considered as such, because the solely size-dependent definitions of the term "nanomaterial" emerged in recent times first in Recommendation 2011/696/EU. Therefore, former physicochemical and toxicological evaluations of SAS were already performed on nanomaterials, however, without being addressed as such. Safety concerns can only emerge if two criteria, (toxicological) hazard and exposure towards the substance is fulfilled at the same time. In case of SAS, the Scientific Committee on Consumer Safety (SCCS) challenged provided data to be insufficient to draw a conclusion regarding the safety of SAS and thus, requested further investigations, in particular by exploring skin penetration of particulate SAS.Investigation of specific particulate substances in skin penetration tests is an analytical challenge. The number of available analytical techniques that are capable to detect nanomaterials in complex matrices, like receptor fluids from skin penetration testing, are limited and still emerging. In the new studies, a comprehensive set of analytical techniques were used to investigate the skin penetration potential of SAS. Particle-sensitive, element and particle-specific combinations of techniques and different sample preparation procedures, that respected the particulate nature of SAS, were used to detect SAS in receptor fluids directly. In addition, electron microscopic techniques were used to examine different layers of skin to detect adsorbed SAS.The combination of Asymmetric Flow Field-Flow Fractionation (AF4) in combination with Inductively Coupled Plasma Mass Spectrometry (ICP-MS) for examination of receptor fluids and Scanning Electron Microscopy coupled with Energy Dispersive X-ray spectroscopy (SEM/EDX) for examination of skin itself, were identified as suitable techniques for the detection of SAS in skin penetration tests. Data from literature was used to compare the results of the studies with the outcome of other test systems (other particles, other techniques). Both, the test results, and literature evaluation led to the conclusion, that SAS does not penetrate skin. Based on this outcome and local and systemic dermal toxicity review of SAS, it can be concluded that dermal application of SAS in cosmetic formulations is negligible. • New analytical systems to detect particles in report fluids are presented. • Synthetic amorphous particles do not penetrate through pig and human skin. • A critical literature review is provided to understand different results reported. [ABSTRACT FROM AUTHOR]

Dr. Helmut Cölfen, an exceptional interdisciplinary scientist, mentor, colleague, and dear friend, passed away in November 2023 at the age of 58. His untimely departure is a profound loss for the fields of analytical ultracentrifugation, colloid, crystallization, and polymer research. This obituary pays tribute to Helmut, honoring his remarkable academic career and contributions to the study of nanochemistry, biophysics, and life sciences. Helmut was renowned for his pioneering research contributions in several key research areas: (1) Development of advanced analytical techniques: Helmut made major contributions to techniques such as analytical ultracentrifugation and field flow fractionation, which are widely utilized to characterize the structure of biomolecules and the growth of nanostructured crystalline materials; (2) Study of nucleation and crystallization processes: Helmut explored the early stages of crystallization which led to the discovery of pre-nucleation clusters and mesocrystal intermediates, in the presence of additives and templates; and (3) Investigation of structure and morphogenesis of mesocrystals, examining their molecular properties. [ABSTRACT FROM AUTHOR]

The use of titanium dioxide (TiO2) as an additive in food has been a subject of ongoing debate in Europe due to its potential toxicity. To inform and support toxicological assessment of this additive, a better understanding of the physicochemical characteristics of TiO2 added to food is needed. For this reason, recent research efforts have focused on method development for the physicochemical characterisation of TiO2 in food and, as a raw material, in a regulatory context. Despite the significant progress made so far, more information is needed about the size-resolved quantification of titania nanoparticles in the fraction below 100 nm. Here, we report the development of a multi-method analytical platform for the characterisation of TiO2 in food. It is based on single particle ICP-MS (spICP-MS), supported by asymmetrical flow field-flow fractionation (AF4) coupled on-line to ICP-MS and multi-angle light scattering (MALS). spICP-MS was optimised to detect TiO2 with sizes down to 15 nm. This enabled us to detect and quantify a fraction of TiO2 particles with sizes below 35 nm, which has not been reported in the literature so far. The presence of this size fraction was confirmed by AF4/MALS/ICP-MS in the raw E171 additive. The relative percentage of particles in this fraction (from the total particle number detected) ranged from 48 to 61% in a range of food products purchased from a local grocery store, with a number concentration range of 2.9 1010–7.13 1013 kg−1, depending on the product. These findings will help bridge the gap with regard to lack of data comparability between orthogonal measurement methods and provide further scientific evidence supporting policy makers and regulators. [ABSTRACT FROM AUTHOR]

Lipid conjugates have advanced the field of lipid-based nanomedicine by promoting active-targeting (ligand, peptide, antibody), stability (PEGylation), controlled release (lipoid prodrug), and probe-based tracking (fluorophore). Recent findings indicate lipid conjugates dissociating from nanomedicine upon encountering a biological environment. Yet, implications for (pre)clinical outcomes remain unclear. In this study, using the zebrafish model (Danio rerio), we investigated the fate of liposome-incorporated lipid fluorophore conjugates (LFCs) after intravenous (IV) administration. LFCs having a bilayer mismatch and relatively polar fluorophore revealed counter-predictive outcomes for Caelyx/Doxil (clearance vs. circulating) and AmBisome-like liposomes (scavenger endothelial cell vs. macrophage uptake). Findings on LFC (mis)match for Caelyx/Doxil-like liposomes were supported by translational intravital imaging studies in mice. Importantly, contradicting observations suggest to originate from LFC dissociation in vivo , which was investigated by Asymmetric Flow Field-Flow Fractionation (AF4) upon liposome-serum incubation in situ. Our data suggests that LFCs matching with the liposome bilayer composition – that did not dissociate upon serum incubation – revealed improved predictive outcomes for liposome biodistribution profiles. Altogether, this study highlights the critical importance of fatty acid tail length and headgroup moiety when selecting lipid conjugates for lipid-based nanomedicine. [Display omitted] [ABSTRACT FROM AUTHOR]

Extracellular vesicles (EVs) have emerged as a promising tool for clinical liquid biopsy. However, the identification of EVs derived from blood samples is hindered by the presence of abundant plasma proteins, which impairs the downstream biochemical analysis of EV‐associated proteins and nucleic acids. Here, we employed optimized asymmetric flow field‐flow fractionation (AF4) combined with density cushion ultracentrifugation (UC) to obtain high‐purity and intact EVs with very low lipoprotein contamination from human plasma and serum. Further proteomic analysis revealed more than 1000 EV‐associated proteins, a large proportion of which has not been previously reported. Specifically, we found that cell‐line‐derived EV markers are incompatible with the identification of plasma‐EVs and proposed that the proteins MYCT1, TSPAN14, MPIG6B and MYADM, as well as the traditional EV markers CD63 and CD147, are plasma‐EV markers. Benefiting from the high‐purity of EVs, we conducted comprehensive miRNA profiling of plasma EVs and nanosized particles (NPs), as well as compared plasma‐ and serum‐derived EVs, which provides a valuable resource for the EV research community. Overall, our findings provide a comprehensive assessment of human blood EVs as a basis for clinical biopsy applications. [ABSTRACT FROM AUTHOR]

The document is a correction notice for an article published in Analytical & Bioanalytical Chemistry. The correction addresses an error in Eq. 29 and the text preceding it. The corrected equations involve ratios of mean fluid velocities and volumetric flow rates in the context of flow field-flow fractionation. The corrections also include adjustments to numerical values quoted in the manuscript. The errors were found to be small under constant channel outlet flow rates, but could become significant if the outlet flow rate varies with time. The corrections have been made to ensure accuracy in the published figures and data tables. [Extracted from the article]

Background The increase in antibiotic resistance is a major public health issue. The development of rapid antimicrobial susceptibility testing (AST) methods is becoming a priority to ensure early and appropriate antibiotic therapy. Objectives To evaluate sedimentation field-flow fractionation (SdFFF) as a method for performing AST in less than 3 h. Methods SdFFF is based on the detection of early biophysical changes in bacteria, using a chromatographic-type technology. One hundred clinical Escherichia coli strains were studied. A calibrated bacterial suspension was incubated for 2 h at 37°C in the absence (untreated) or presence (treated) of five antibiotics used at EUCAST breakpoint concentrations. Bacterial suspensions were then injected into the SdFFF machine. For each E. coli isolate, retention times and elution profiles of antibiotic-treated bacteria were compared with retention times and elution profiles of untreated bacteria. Algorithms comparing retention times and elution profiles were used to determine if the strain was susceptible or resistant. Performance evaluation was done according to CLSI and the ISO standard 20776-2:2021 with broth microdilution used as the reference method. Results AST results from SdFFF were obtained in less than 3 h. SdFFF showed high categorical agreement (99.8%), sensitivity (99.5%) and specificity (100.0%) with broth microdilution. Results for each antimicrobial were also in agreement with the ISO 20776-2 recommendations, with sensitivity and specificity of ≥95.0%. Conclusions This study showed that SdFFF can be used as a rapid, accurate and reliable phenotypic AST method with a turnaround time of less than 3 h. [ABSTRACT FROM AUTHOR]

Recently, the first generic glucagon for injection was approved for the treatment of severe hypoglycemia. Unlike its brand name recombinant glucagon, the generic glucagon is synthetic. Since glucagon has a high propensity to form aggregates in solution, it is essential to assess the aggregation profile of the synthetic glucagon compared to the recombinant glucagon. In this study, two robust separation methods, size-exclusion chromatography (SEC-HPLC) and field-flow fractionation coupled with a multi-angle light scattering detector (FFF-MALS), were employed to characterize generic and brand glucagon aggregation in six lots (three newly released, three expired). The presence of aggregation in samples was determined from the generated chromatograms and analyzed. The study showed that both products have comparable aggregation profiles. The SEC-HPLC demonstrated that in both glucagon versions, the expired lots had a higher percentage of dimers than the newly released lots, but even at expiration, the amount was negligible (∼0.1%). The FFF-MALS method did not detect any dimers or higher molecular weight aggregates. Further evaluation of the detection limit found that FFF-MALS was unable to detect aggregates at amounts lower than 0.5% of total glucagon. The negligible amounts of dimer detected in the generic and brand glucagon indicate that both versions are physically stable and are not prone to aggregation under clinically relevant conditions. [ABSTRACT FROM AUTHOR]

The article "Reply to: Technical challenges of studying the impact of plasma components on the efficacy of lipid nanoparticles for vaccine and therapeutic applications" discusses the findings of a study on the function of therapeutic lipid nanoparticles (LNPs) and their association with plasma components. The study found that LNPs with higher efficacy tended to have increased high-density lipoprotein (HDL) content. The authors used a pull-down method to isolate LNPs from plasma and conducted various experiments to validate their findings. They addressed concerns about potential contamination and emphasized the need for careful assay design and validation when studying LNPs and other nanosized particles. [Extracted from the article]

Colloids can facilitate the transport of trace elements in soil, but earlier studies fell short either because of sampling artefacts or because no contrasting soils were used; both factors hamper a better understanding of where this occurs. Here, leaching in undisturbed and unsaturated soil columns was observed to identify colloidal transport of cadmium (Cd) and uranium (U) in soils with contrasting properties. Crucially, it was examined how such column data relate to more traditional in situ data obtained with porous suction cups. Soils were collected from three agricultural fields in Switzerland (soil pH 5.1–7.3), leached during 36 days (0.6 pore volumes, ~1.6 mm day−1), and elements trapped in the PTFE suction plates were recovered and were ascribed to colloids. Of the total mobile mass (sum percolate and plate), 18%–53% (Cd) or 8–89% (U) was retained by plates, the range depending on the field where soil was sampled and that fraction depends on soil pH and for U also on dissolved inorganic carbon. The total mobile Cd or U mass divided by the percolated volume indicates the metal concentration in the average leachate and was logically highest for Cd in the most acidic soil and for U in the most alkaline soil. The concentrations of Cd and U in these column leachates were >10‐fold higher than corresponding concentrations in the suction cups in the field for those soils and elements where colloids dominated leaching. In contrast, they were within a factor of two where colloids are <20% of the mobile mass. Flow Field‐Flow Fractionation (FlFFF) analyses showed an association of colloidal U with oxyhydroxides, clays and organic matter and, for colloidal Cd, an association with organic matter. These data show that suction cups or filtered percolates can underestimate the leaching of Cd and U by up to an order of magnitude. [ABSTRACT FROM AUTHOR]

We established a size separation method for silica nanoparticles (SiNPs) measuring 10, 30, 50, 70, and 100 nm in diameter using asymmetric flow field flow fractionation hyphenated with inductively coupled plasma mass spectrometry (AF4-ICP-MS), and evaluated the cytotoxicity of SiNPs in human hepatoma HepG2 cells. Analysis of the mixture sample revealed that nanoparticles of different sizes were eluted at approximately 2-min intervals, with no effect on each elution time or percentage recovery. Compared with larger SiNPs, smaller SiNPs exhibited high cytotoxicity when the volume of SiNPs exposed to the cells was the same. We measured SiNPs in culture medium and inside cells by AF4-ICP-MS and found that approximately 17% of SiNPs in the mixture of five differently sized particles were absorbed by the cells. Transmission electron microscopy revealed that 10 nm SiNPs formed aggregates and accumulated in the cells. Based on AF4-ICP-MS analysis, there is no clear difference in the particle volume absorbed by the cells among different sizes. Therefore, the high toxicity of small SiNPs can be explained by the fact that their large surface area relative to particle volume efficiently induces toxicological influences. Indeed, the large surface area of 10 nm SiNPs significantly contributed to the production of reactive oxygen species. [ABSTRACT FROM AUTHOR]

The co-transport of contaminants by soil colloids can generate substantial environmental risk, and this behavior is greatly affected by environmental conditions. In this study, AF4-ICP-MS was used to investigate the size distribution and composition of Cd/Pb-bearing colloids; saturated sand column experiments were used to investigate the impact of soil colloids on the transport of Cd/Pb under different pH and ionic strength conditions. AF4-ICP-MS characterization showed that natural colloids were primarily associated with two sizes ranges: 0.3–35 KDa (F1, fine nanoparticles) and 280 KDa–450 nm (F2, larger nanoparticles), which mainly consisted of organic matter (OM), iron (Fe), and manganese (Mn) (oxy)hydroxides and clay minerals. Fine nanoparticles could strongly adsorb Cd and Pb under all environmental conditions. Mn and Fe (oxy)hydroxides generally formed under neutral to alkaline conditions and exhibited adsorption capabilities for Cd and Pb, respectively. Transport experiments were conducted under different pH and ionic strength conditions. At pH 3.0, soil colloids had little effect on the transport of Cd2+ and Pb2+. At pH 5.0, soil colloids inhibited the transport of Cd2+ by 16.1%, and Pb2+ recovery was still 0.0%. At pH 7.0 and 9.0, soil colloids facilitated the transport of Cd2+ by 15.6% and 29.6%, facilitated Pb2+ by 1.3% and 6.4%. At an ionic strength of 0, 0.005, and 0.01 mol L−1 NaNO3, soil colloids facilitated the transport of Cd2+ by 77.7%, 45.8%, and 15.6%, only facilitated the transport of Pb2+ by 46.2% at an ionic strength of 0 mol L−1 NaNO3. At an ionic strength of 0.05 mol L−1 NaNO3, soil colloids inhibited the transport of Cd2+ and Pb2+ by 33.1% and 21.0%, respectively. The transport of Cd2+ and Pb2+ facilitated by soil colloids was clearly observed under low ionic strength and non-acidic conditions, which can generate a potential environmental risk. [ABSTRACT FROM AUTHOR]

Palate fullness (PF) and mouthfeel are important sensory attributes influencing beer quality. The molar mass of starch (dextrins) and non-starch (β-glucans) polysaccharides may influence PF (pleasant) or mouthfeel (sliminess, unpleasant), respectively. Therefore, this research aims to generate beer with wide physico-chemical responses based on various raw material characteristics to study its relation to PF and mouthfeel. To accomplish this, ten barley varieties (two harvest locations and years) were classified into three groups based on their modification characteristics. To intensify response variation, barley was malted at two modification levels (parameter steeping degree), generating 55 independent malts used to brew the same number of standardized bottom fermented beers. A trained sensory panel evaluated PF (intensity and quality) and mouthfeel (e.g., slimy) descriptors. Additionally, beers were fractionated by asymmetrical flow field-flow fractionation (AF4) in three different fractions and their molar masses were determined. The average molar mass of big size (> 10 nm) dextrins and β-glucans AF4 fraction increased analogously to barley modification characteristics. For sensory data evaluation, only beer samples brewed with malts inside the recommended brewing specifications were considered (β-glucan content in malt < 350 mg/L, ISO 65 °C). PF quality was lower on samples with β-glucan content < 100 mg/L. This group also contained fewer dextrins (attenuation > 87.4 ± 1.5%), suggesting that the balance of β-glucans and dextrins is important for PF quality. This work indicates that molar mass of starch and non-starch polysaccharides, affected by barley variety and its modification level, influences sensory perception, and hence, beer quality. [ABSTRACT FROM AUTHOR]

The EU has approved the usage of gold as a food additive (E175) and it has been applied in numerous foods for coloring and decoration purposes. Different from the general assumption that edible gold is mainly present in the form of flakes or external coating in foods, this work demonstrated that gold nanoparticles (Au NPs) can be released from gold flakes and extracted under optimized conditions. To support future risk assessment associated with the exposure of Au NPs to human health, an effective approach was established in this study for both size characterization and mass determination of Au NPs released in a commercial gold-containing liquor using Asymmetric Flow Field-flow Fractionation (AF4) hyphenated with Inductively Coupled Plasma Mass Spectrometry (ICP-MS). Our results showed that no Au NPs were detected in the original liquor product and only after ultrasonication for several minutes did Au NPs occur in the ultrasound-treated liquor. Particularly, Au NPs released in the liquor can be well extracted after 100-fold enrichment of gold flakes and the subsequent ultrasonication for 25 min. Size characterization of Au NPs was conducted by AF4-ICP-MS under calibration with Au NP standards. The gold particle sizes detected ranged from 8.3–398.0 nm and the dominant size of the released Au NPs was around 123.7 nm in the processed liquor. The mass concentration of gold particles determined in the liquor sample with gold flakes concentrated and subsequently sonicated was 48.1 μg L−1 by pre-channel calibration and the overall detection recoveries ranged over 82–95%. For the comparison control samples without ultrasonication, there was no detection of Au NPs. The established method was demonstrated to be useful for monitoring Au NPs in liquor and is possibly applied to other similar foodstuffs. [ABSTRACT FROM AUTHOR]

Copper (Cu) has been used to treat vines for a long time, which has led to its accumulation in vineyard soils. In the present work, the mobilization of copper from these soils and its transport, and diffusion outside the plots by drain water were investigated. For this, the distribution of copper between the dissolved and colloidal phases, and within the colloidal phase, of these waters was determined using an investigation strategy based on the coupling between a size separation technique, asymmetric flow field-flow fractionation, and several detectors. First, the total copper concentrations in water from different drains were monitored over a period of 2 years: Cu was mainly found in the fraction of < 450 nm. Then, the distribution of copper on the size continuum was more closely studied in water from one of the drains, sampled over a winter period. Between 45 and 75% of Cu was found in the 2–450 nm colloidal fraction. The <450 nm colloidal phase of the drain waters was found to be mainly composed of humic acids (~15 to 60 mg L−1) and clay-rich particles (~100 to 650 mg (Al) L−1). These particles also contained (hydr)oxides of iron and manganese. The concentrations of Fe and Mn were approximately 100 to 200 times lower than those of Al. The majority of humic acids had an apparent molar mass of ≤ 10 kDa. They were distributed along the size continuum: (i) in a population with an average size of ~20 nm, probably consisting of supramolecular entities, and (ii) associated with clay-rich particles with a size of ~120–200 nm. Copper was found to be complexed with humic acids and associated with clays via clay-humic complexes. Copper mobilization from the soil to the water and its transport to the drain water appeared governed by the soil humidity level and the rainfall. [ABSTRACT FROM AUTHOR]

The early-stage diagnosis of cancer is a crucial clinical need. The inadequacies of surgery tissue biopsy have prompted a transition to a less invasive profiling of molecular biomarkers from biofluids, known as liquid biopsy. Exosomes are phospholipid bilayer vesicles present in many biofluids with a biologically active cargo, being responsible for cell-to-cell communication in biological systems. An increase in their excretion and changes in their cargo are potential diagnostic biomarkers for an array of diseases, including cancer, and they constitute a promising analyte for liquid biopsy. The number of exosomes released, the morphological properties, the membrane composition, and their content are highly related to the physiological and pathological states. The main analytical challenge to establishing liquid biopsy in clinical practice is the development of biosensors able to detect intact exosomes concentration and simultaneously analyze specific membrane biomarkers and those contained in their cargo. Before analysis, exosomes also need to be isolated from biological fluids. Microfluidic systems can address several issues present in conventional methods (i.e., ultracentrifugation, size-exclusion chromatography, ultrafiltration, and immunoaffinity capture), which are time-consuming and require a relatively high amount of sample; in addition, they can be easily integrated with biosensing systems. A critical review of emerging microfluidic-based devices for integrated biosensing approaches and following the major analytical need for accurate diagnostics is presented here. The design of a new miniaturized biosensing system is also reported. A device based on hollow-fiber flow field-flow fractionation followed by luminescence-based immunoassay is applied to isolate intact exosomes and characterize their cargo as a proof of concept for colon cancer diagnosis. [ABSTRACT FROM AUTHOR]

[Display omitted] • Minor structural differences of hydrophobic porphyrins can distinctly influence their release properties from liposomes. • mTHPP (meta OH) redistributes much slower than pTHPP (para OH) from liposomal membranes. • Reversed-phase HPLC experiments indicate a higher partition coefficient for mTHPP. • In MD simulations, mTHPP has a higher tendency for hydrogen bonding, a larger tilt angle and locates deeper in the membrane. • Both HPLC and MD simulation results support the observed differences in release and redistribution of mTHPP and pTHPP from liposomal membranes. The release of hydrophobic compounds from liposomal membranes occurs by partitioning and is thus determined by the physicochemical properties (e.g. logP and water solubility) of the drug. We postulate that even minor structural differences, e.g. the position of the phenolic OH-group of the hydrophobic porphyrins mTHPP and pTHPP (meta vs. para substitution), distinctly affect their partitioning and release behavior from liposomes. The release and redistribution of mTHPP and pTHPP from lecithin or POPC/POPG liposomes to different acceptor particles (DSPE-mPEG micelles and liposomes) was studied by asymmetrical flow field-flow fractionation to separate donor and acceptor particles. Reversed phase HPLC was applied to detect differences in partitioning. Molecular dynamics (MD) simulations were carried out to obtain molecular insight in the different behavior of the two compounds inside a lipid bilayer. Despite the minor differences in chemical structure, mTHPP is more hydrophobic and redistributes much slower to both acceptor phases than pTHPP. MD simulations indicate that compared to pTHPP, mTHPP makes stronger hydrogen bonds with the lipid head groups, is oriented more parallel to the lipid tails and is embedded slightly deeper in the membrane. [ABSTRACT FROM AUTHOR]

We present a generically applicable approach to determine an extensive set of size-dependent critical quality attributes inside nanoparticulate pharmaceutical products. By coupling asymmetrical-flow field-flow fractionation (AF4) measurements directly in-line with solution small angle X-ray scattering (SAXS), vital information such as (i) quantitative, absolute size distribution profiles, (ii) drug loading, (iii) size-dependent internal structures, and (iv) quantitative information on free drug is obtained. Here the validity of the method was demonstrated by characterizing complex mRNA-based lipid nanoparticle products. The approach is particularly applicable to particles in the size range of 100 nm and below, which is highly relevant for pharmaceutical products—both biologics and nanoparticles. The method can be applied as well in other fields, including structural biology and environmental sciences. [ABSTRACT FROM AUTHOR]

Acrylated methyl esters of higher fatty acids derived from camelina oil and linseed oil were synthesized through transesterification, epoxidation, and subsequent acrylation. Methyl methacrylate and butyl acrylate were copolymerized with various amounts of bio-based derivatives (5 – 30 wt% in monomer mixture) to obtain polymeric latexes for coating applications. Successful emulsion polymerizations with up to 25 wt% of the bio-based derivatives were performed with low coagulum (below 2%) and high monomer conversion (around 95%). The incorporation of bio-based derivatives into polymeric latexes was confirmed with infrared spectroscopy. Asymmetric flow field flow fractionation coupled with a multi-angle light scattering was used to analyze the synthesized copolymers in terms of their molar mass distribution. The results revealed that copolymerizing the bio-based derivatives resulted in ultra-high molar mass nanogel fractions formed because of multi-acrylated ingredients derived from polyunsaturated fatty acids. The phenomenon of nanogel formation became more pronounced for the linseed oil-based derivative. Evaluated coating properties showed that latexes comprising the bio-based derivatives provided increased water repellence (about 10° higher water contact angles were achieved for all bio-based coating compositions in contrast to a reference latex). Moreover, latexes comprising chemically modified oils in the content of 25 and 30 wt% provided water whitening-resistant coatings, making the bio-based derivatives promising candidates for replacing petroleum-based monomers in the production of sustainable latex coatings. [ABSTRACT FROM AUTHOR]

Nanohydrogel particles of polyethylene glycol (PEG), gelatin (GEL), and PEG–GEL mixtures (MIXs) were synthesized with a high electron beam and 60Co gamma-ray radiation. The relatively novel technique of Asymmetrical Flow Field Flow Fractionation (AF4 or AFFFF) coupled to a Multi-Angle Laser Light Scattering (MALLS) detector was mainly used to determine the hydrodynamic diameter (Dh) of the radiation-synthesized PEG, GEL, and PEG–GEL nanohydrogel particles. Our approach to achieving nanohydrogel particles is to enhance the intracrosslinking reactions and decrease the intercrosslinking reactions of the C-centered radicals of the PEG and GEL. The intracrosslinking reactions of these free radicals were enhanced via irradiation at temperatures of 77–80 °C and using a high dose rate and pulsed irradiation. The shorter average distance between the C-centered free radicals on the backbone of the thermally collapsed PEG and GEL chain, due to the destruction of hydrogen bonds, enhances the intracrosslinking reactions. It was observed that increasing the dose and dose rate decreased the Dh. DLS results lined up with AF4 measurements. This study provides researchers with a clean method to produce GEL–PEG hydrogels without the use of toxic reagents. Particle size can be tuned with dose, dose rate, and temperature as demonstrated in this work. This is ideal for medical applications as the use of ionizing radiation eliminates toxicity concerns and provides simultaneous sterilization of the material. [ABSTRACT FROM AUTHOR]

Coiled tube field-flow fractionation (CTFFF) is currently applied to environmental and material studies. In the present work, a novel zone elution mode in CTFFF has been proposed and developed. Zone elution mode is based on the separation of particles by stepwise decreasing the flow rate of the carrier fluid and their subsequent elution at a constant flow rate. The fractionation parameters were optimized using a mixture of standard silica submicron particles (150, 390, and 900 nm). Taking samples of volcanic ash as examples, it has been demonstrated that zone elution mode can be successfully used for the fractionation of environmental nano- and submicron particles. For the first time, CTFFF was coupled online with a dynamic light scattering detector for the size characterization of eluted particles. Zone elution in CTFFF can serve for the further development of hyphenated techniques enabling efficient fractionation and size/elemental characterization of environmental particles in nano- and submicrometric size ranges. [ABSTRACT FROM AUTHOR]

Field-flow fractionation (FFF) is a family of single-phase separative techniques exploited to gently separate and characterize nano- and microsystems in suspension. These techniques cover an extremely wide dynamic range and are able to separate analytes in an interval between a few nm to 100 µm size-wise (over 15 orders of magnitude mass-wise). They are flexible in terms of mobile phase and can separate the analytes in native conditions, preserving their original structures/properties as much as possible. Molecular biology is the branch of biology that studies the molecular basis of biological activity, while biotechnology deals with the technological applications of biology. The areas where biotechnologies are required include industrial, agri-food, environmental, and pharmaceutical. Many species of biological interest belong to the operational range of FFF techniques, and their application to the analysis of such samples has steadily grown in the last 30 years. This work aims to summarize the main features, milestones, and results provided by the application of FFF in the field of molecular biology and biotechnology, with a focus on the years from 2000 to 2022. After a theoretical background overview of FFF and its methodologies, the results are reported based on the nature of the samples analyzed. [ABSTRACT FROM AUTHOR]

Liposome development is of great interest owing to increasing requirements for efficient drug carriers. The structural features and thermal stability of such liposomes are crucial in drug transport and delivery. Reported here are the results of the structural characterization of PEGylated liposomes via small‐ and wide‐angle X‐ray scattering and an asymmetric flow field‐flow fractionation (AF4) system coupled with differential refractive‐index detection, multi‐angle light scattering (MALS) and dynamic light scattering. This integrated analysis of the exemplar PEGylated liposome formed from hydrogenated soy phosphatidylcholine (HSPC) with the addition of cholesterol reveals an average hydrodynamic radius (Rh) of 52 nm with 10% polydispersity, a comparable radius of gyration (Rg) and a major liposome particle mass of 118 kDa. The local bilayer structure of the liposome is found to have asymmetric electronic density profiles in the inner and outer leaflets, sandwiched by two PEGylated outer layers ca 5 nm thick. Cholesterol was found to effectively intervene in lipid chain packing, resulting in the thickening of the liposome bilayer, an increase in the area per lipid and an increase in liposome size, especially in the fluid phase of the liposome. These cholesterol effects show signs of saturation at cholesterol concentrations above ca 1:5 cholesterol:lipid molar ratio. [ABSTRACT FROM AUTHOR]

Characterization of particulate impurities such as aggregates is necessary to develop safe and efficacious adeno-associated virus (AAV) drug products. Although aggregation of AAVs can reduce the bioavailability of the virus, only a limited number of studies focus on the analysis of aggregates. We explored three technologies for their capability to characterize AAV monomers and aggregates in the submicron (<1 µm) size range: (i) mass photometry (MP), (ii) asymmetric flow field flow fractionation coupled to a UV-detector (AF4-UV/Vis) and (iii) microfluidic resistive pulse sensing (MRPS). Although low counts for aggregates impeded a quantitative analysis, MP was affirmed as an accurate and rapid method for quantifying the genome content of empty/filled/double-filled capsids, consistent with sedimentation velocity analytical ultracentrifugation results. MRPS and AF4-UV/Vis enabled the detection and quantification of aggregate content. The developed AF4-UV/Vis method separated AAV monomers from smaller aggregates, thereby enabling a quantification of aggregates <200 nm. MRPS was experienced as a straightforward method to determine the particle concentration and size distribution between 250-2000 nm, provided that the samples do not block the microfluidic cartridge. Overall, within this study we explored the benefits and limitations of the complementary technologies for assessing aggregate content in AAV samples. [ABSTRACT FROM AUTHOR]

Proliferating cell nuclear antigen (PCNA) is the key regulator of human DNA metabolism. One important interaction partner is p15, involved in DNA replication and repair. Targeting the PCNA–p15 interaction is a promising therapeutic strategy against cancer. Here, a Förster resonance energy transfer (FRET)-based assay for the analysis of the PCNA–p15 interaction was developed. Next to the application as screening tool for the identification and characterization of PCNA–p15 interaction inhibitors, the assay is also suitable for the investigation of mutation-induced changes in their affinity. This is particularly useful for analyzing disease associated PCNA or p15 variants at the molecular level. Recently, the PCNA variant C148S has been associated with Ataxia-telangiectasia-like disorder type 2 (ATLD2). ATLD2 is a neurodegenerative disease based on defects in DNA repair due to an impaired PCNA. Incubation time dependent FRET measurements indicated no effect on PCNAC148S–p15 affinity, but on PCNA stability. The impaired stability and increased aggregation behavior of PCNAC148S was confirmed by intrinsic tryptophan fluorescence, differential scanning fluorimetry (DSF) and asymmetrical flow field-flow fractionation (AF4) measurements. The analysis of the disease associated PCNA variant demonstrated the versatility of the interaction assay as developed. [ABSTRACT FROM AUTHOR]

Nanogels are candidates for biomedical applications, and core-shell nanogels offer the potential to tune thermoresponsive behaviour with the capacity for extensive degradation. These properties were achieved by the combination of a core of poly(N-isopropylmethacrylamide) and a shell of poly(N-isopropylacrylamide), both crosslinked with the degradable crosslinker N,N′-bis(acryloyl)cystamine. In this work, the degradation behaviour of these nanogels was characterised using asymmetric flow field flow fractionation coupled with multi-angle and dynamic light scattering. By monitoring the degradation products of the nanogels in real-time, it was possible to identify three distinct stages of degradation: nanogel swelling, nanogel fragmentation, and nanogel fragment degradation. The results indicate that the core-shell nanogels degrade slower than their non-core-shell counterparts, possibly due to a higher degree of self-crosslinking reactions occurring in the shell. The majority of the degradation products had molecule weights below 10 kDa, which suggests that they may be cleared through the kidneys. This study provides important insights into the design and characterisation of degradable nanogels for biomedical applications, highlighting the need for accurate characterisation techniques to measure the potential biological impact of nanogel degradation products. [ABSTRACT FROM AUTHOR]

The fusion and classification of hyperspectral images (HSIs) and light detection and ranging (LiDAR) data have been extensively studied using deep learning. However, traditional real-valued deep-learning methods have limitations in distinguishing internal and external relations and capturing fine spatial characteristics. To break through these limitations, this letter proposes a quaternion convolutional neural network (QCNN) with extended morphological attribute profile (EMAP) quaternion representation (called EQR) for multisource remote-sensing (RS) data classification by utilizing quaternion properties. Specifically, we first propose the EQR for each single-source data, which encodes the multiattribute features in a compact, yet comprehensive manner, highlighting the internal relations. Second, we embed EQR into QCNN to preserve the internal relations and enable the interaction of multiattribute features. Then, we develop the 3-D quaternion convolution (3DQConv) to better exploit the 3-D characteristic of HSIs. Finally, we design different attention mechanisms and a two-level fusion strategy for multisource data to learn enhanced features. Experiments on two multisource RS datasets show that the proposed method achieved better performance than other state-of-the-art classification methods. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Extractions methods based on ultrapure water, tetramethylammonium hydroxide (TMAH), and tetrasodium pyrophosphate (TSPP) were applied to faeces collected from two in vivo experiments of pigs and chickens fed with a silver-based nanomaterial to study the fate and speciation of silver. For TMAH extraction, cysteine and CaCl2 were used to evaluate their stabilization effect on the silver forms. The analytical techniques single-particle inductively coupled plasma mass spectrometry (SP-ICP-MS), hydrodynamic chromatography hyphenated to ICP-MS (HDC-ICP-MS) and asymmetric flow field flow fractionation coupled to ICP-MS (AF4-ICP-MS) were applied to the simultaneous detection of particulate and dissolved silver. Results have shown that water extraction was a suitable option to assess the environmental release of silver, with percentages of 3 and 9% for faeces of pigs and chickens, respectively. The use of TMAH extraction combined with SP-ICP-MS analysis was useful to characterize Ag-containing particles (less than 1%). Both stabilizers, cysteine and CaCl2, have a similar effect on silver nanoparticle preservation for chicken faeces, whereas cysteine-Triton was better for pig samples. In any case, silver extraction efficiency with TMAH was low (39–42%) for both types of faeces due to a matrix effect. TSPP followed by ICP-MS enabled the fractionation of the silver in the faeces, with silver sulphide (41%) and ionic silver (62%) being the most abundant fractions. [ABSTRACT FROM AUTHOR]

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Asymmetric-flow field-flow fractionation (AF4) is a gentle, flexible, and powerful separation technique that is widely utilized for fractionating nanometer-sized analytes, which extend to many emerging nanocarriers for drug delivery, including lipid-, virus-, and polymer-based nanoparticles. To ascertain quality attributes and suitability of these nanostructures as drug delivery systems, including particle size distributions, shape, morphology, composition, and stability, it is imperative that comprehensive analytical tools be used to characterize the native properties of these nanoparticles. The capacity for AF4 to be readily coupled to multiple online detectors (MD-AF4) or non-destructively fractionated and analyzed offline make this technique broadly compatible with a multitude of characterization strategies, which can provide insight on size, mass, shape, dispersity, and many other critical quality attributes. This review will critically investigate MD-AF4 reports for characterizing nanoparticles in drug delivery, especially those reported in the last 10–15 years that characterize multiple attributes simultaneously downstream from fractionation. [ABSTRACT FROM AUTHOR]

Carbon black nanomaterial (CB-NM), as an industrial product with a large number of applications, poses a high risk of exposure, and its impact on health needs to be assessed. The most common testing platform for engineered (E)NMs is in vitro toxicity assessment, which requires prior ENM dispersion, stabilization, and characterization in cell culture media. Here, asymmetric flow field-flow fractionation (AF4) coupled to UV–Vis and dynamic light scattering (DLS) detectors in series was used for the study of CB dispersions in cell culture media, optimizing instrumental variables and working conditions. It was possible to disperse CB in a non-ionic surfactant aqueous solution due to the steric effect provided by surfactant molecules attached on the CB surface which prevented agglomeration. The protection provided by the surfactant or by culture media alone was insufficient to ensure good dispersion stability needed for carrying out in vitro toxicity studies. On the other hand, cell culture media in combination with the surfactant improved dispersion stability considerably, enabling the generation of shorter particles and a more favourable zeta potential magnitude, leading to greater stability due to electrostatic repulsion. It was demonstrated that the presence of amino acids in the culture media improved the monodisperse nature and stability of the CB dispersions, and resulted in a turn towards more negative zeta potential values when the pH was above the amino acid isoelectric point (IEP). Culture media used in real cell culture scenarios were also tested, and in vitro toxicity assays were developed optimizing the compatible amount of surfactant. [ABSTRACT FROM AUTHOR]

Electrical asymmetric-flow field-flow fractionation (EAF4) is a new and interesting analytical technique recently proposed for the characterization of metallic nanoparticles (NPs). It has the potential to simultaneously provide relevant information about size and electrical parameters, such as electrophoretic mobility (μ) and zeta-potential (ζ), of individual NP populations in an online instrumental setup with an array of detectors. However, several chemical and instrumental conditions involved in this technique are definitely influential, and only few applications have been proposed until now. In the present work, an EAF4 system has been used with different detectors, ultraviolet–visible (UV–vis), multi-angle light scattering (MALS), and inductively coupled plasma with triple quadrupole mass spectrometry (ICP-TQ-MS) for the characterization of gold, silver, and platinum NPs with both citrate and phosphate coatings. The behavior of NPs has been studied in terms of retention time and signal intensity under both positive and negative current with results depending on the coating. Carrier composition, particularly ionic strength, was found to be critical to achieve satisfactory recoveries and a reliable measurement of electrical parameters. Dynamic light scattering (DLS) has been used as a comparative technique for these parameters. The NovaChem surfactant mix (0.01%) showed a quantitative recovery (93 ± 1%) of the membrane, but the carrier had to be modified by increasing the ionic strength with 200 μM of Na2CO3 to achieve consistent μ values. However, ζ was one order of magnitude lower in EAF4-UV–vis-MALS than in DLS, probably due to different electric processes in the channel. From a practical point of view, EAF4 technique is still in its infancy and further studies are necessary for a robust implementation in the characterization of NPs. [ABSTRACT FROM AUTHOR]

Chitosan is extensively studied as a carrier for gene delivery and is an attractive non-viral gene vector owing to its polycationic, biodegradable, and biocompatible nature. Thus, it is essential to understand the chemistry of self-assembled chitosan–DNA complexation and their structural and functional properties, enabling the formation of an effective non-viral gene delivery system. In this study, two parent chitosans (samples NAS-032 and NAS-075; Mw range ~118–164 kDa) and their depolymerised derivatives (deploy nas-032 and deploy nas-075; Mw range 6–14 kDa) with degrees of acetylation 43.4 and 4.7%, respectively, were used to form polyelectrolyte complexes (PECs) with DNA at varying [–NH3+]/[–PO4−] (N/P) molar charge ratios. We investigated the formation of the PECs using ζ-potential, asymmetric flow field-flow fractionation (AF4) coupled with multiangle light scattering (MALS), refractive index (RI), ultraviolet (UV) and dynamic light scattering (DLS) detectors, and TEM imaging. PEC formation was confirmed by ζ-potential measurements that shifted from negative to positive values at N/P ratio ~2. The radius of gyration (Rg) was determined for the eluting fractions by AF4-MALS-RI-UV, while the corresponding hydrodynamic radius (Rh), by the DLS data. We studied the influence of different cross-flow rates on AF4 elution patterns for PECs obtained at N/P ratios 5, 10, and 20. The determined rho shape factor (ρ = Rg/Rh) values for the various PECs corresponded with a sphere morphology (ρ ~0.77–0.85), which was consistent with TEM images. The results of this study represent a further step towards the characterisation of chitosan–DNA PECs by the use of multi-detection AF4 as an important tool to fractionate and infer aspects of their morphology. [ABSTRACT FROM AUTHOR]

Extracellular vesicles (EV) have many attributes important for biomedicine; however, current EV isolation methods require long multi-step protocols that generally involve bulky equipment that cannot be easily translated to clinics. Our aim was to design a new cyclic olefin copolymer–off-stoichiometry thiol-ene (COC–OSTE) asymmetric flow field fractionation microfluidic device that could isolate EV from high-volume samples in a simple and efficient manner. We tested the device with large volumes of urine and conditioned cell media samples, and compared it with the two most commonly used EV isolation methods. Our device was able to separate particles by size and buoyancy, and the attained size distribution was significantly smaller than other methods. This would allow for targeting EV size fractions of interest in the future. However, the results were sample dependent, with some samples showing significant improvement over the current EV separation methods. We present a novel design for a COC–OSTE microfluidic device, based on bifurcating asymmetric flow field-flow fractionation (A4F) technology, which is able to isolate EV from large volume samples in a simple, continuous-flow manner. Its potential to be mass-manufactured increases the chances of implementing EV isolation in a clinical or industry-friendly setting, which requires high repeatability and throughput. [ABSTRACT FROM AUTHOR]

After pyrolysis at 600 °C and enrichment of the degradation products on solid-phase absorbers, plastic species were quantified by GC-MS analysis detecting polymer-specific decomposition products [271]. Furthermore, unlike vibrational spectroscopy techniques such as Raman, it is not sensitive to variables such as colors. Raman method spectroscopy (<125 mm and >5 mass%) [266], micro-Raman spectroscopy (>100 µm), micro-FTIR spectroscopy (>100 µm) [267], m-Raman spectroscopy (>1 mm), m-FTIR spectroscopy (>10 mm) [268], macroscopic dimensioned near-infrared (NIR) in combination with chemometrics (>10 mm and 1 mass%) [266], and hyperspectral imaging technology (0.5-5 mm) can be used to identify plastics type, but these techniques are time-consuming, and they can be expensive and result in an uncertain extrapolation of MP quantity [269]. [Extracted from the article]

White blood cell (WBC) count will greatly facilitate the timely diagnosis and management of health conditions of dairy cattle, including clinical mastitis. We propose a WBC counting device for dairy cattle using a microchannel with field-flow fractionation incorporating light scattering, which allows both the separation and counting of WBCs. The performance characteristics of the microchannel were optimized to separate particles by size through three-dimensional (3D) numerical simulations. The fabricated microchannel was evaluated using three mock beads with sizes of 3.5 pm representing milk fat globules (MFGs), 10 pm for WBCs, and 30 pm for cancer cells. The pinched flow microfluidic device was useful for separating particles with diameters of 10 and 30 pm. The WBC count was determined using the fabricated WBC counting device. Light scattering was effective for separating cells of 10 pm diameter or less such as MFGs and WBCs that pass through a close streamline. The linear regression analysis of the calibration curve for the WBC counting device showed a multiple determination coefficient of 0.98. The performance of our WBC counting device approached that of laboratory-based tests, allowing the quantitative and rapid analysis of the concentration of WBCs in raw bovine milk in field applications. [ABSTRACT FROM AUTHOR]

In a previous study we used asymmetric-flow field-flow fractionation to determine the polymer mass (Mw), gyration radius (Rw) and the polydispersity index of glutenin polymers (GPs) in wheat (Triticum aestivum). Here, using the same multi-location trials (4 years, 11 locations, and 192 cultivars), we report the factors that are associated with the conformation (Conf) of the polymers, which is the slope of Log(Rw) versus a function of Log(Mw). We found that Conf varied between 0.285 and 0.740, it had low broad-sense heritability (H 2=16.8), and it was significantly influenced by the temperature occurring over the last month of grain filling. Higher temperatures were found to increase Rw and the compactness and sphericity of GPs. Alleles for both high- and low-molecular-weight glutenin subunits had a significant influence on the Conf value. Assuming a Gaussian distribution for Mw, the number of polymers present in wheat grains was computed for different kernel weights and protein concentrations, and it was found to exceed 1012 GPs per grain. Using atomic force microscopy and cryo-TEM, images of GPs were obtained for the first time. Under higher average temperature, GPs became larger and more spherical and consequently less prone to rapid hydrolysis. We propose some orientations that could be aimed at potentially reducing the impact of numerous GPs on people suffering from non-celiac gluten sensitivity. [ABSTRACT FROM AUTHOR]

To predict the polymer properties produced by free‐radical polymerization of N‐vinylpyrrolidone (NVP) in aqueous solution a detailed kinetic model has been developed. The kinetic model allows to calculate the chain length distribution, the number of branching points, and the number of terminal double bonds (TDB). The latter is accounted for since TDBs are a precondition for branching. While monomer conversion can be predicted sufficiently using independently determined rate constants for propagation and termination, here the predictions of structural properties by a newly developed extended kinetic model to experimental findings are compared. Polymer produced in a continuous stirred tank reactor is analyzed by gel permeation chromatography (GPC), field flow fractionation (FFF), and high‐pressure liquid chromatography (HPLC). [ABSTRACT FROM AUTHOR]

The co-transport of contaminants by soil colloids can generate substantial environmental risk, and this behavior is greatly affected by environmental conditions. In this study, AF4-ICP-MS was used to investigate the size distribution and composition of Cd/Pb-bearing colloids; saturated sand column experiments were used to investigate the impact of soil colloids on the transport of Cd/Pb under different pH and ionic strength conditions. AF4-ICP-MS characterization showed that natural colloids were primarily associated with two sizes ranges: 0.3–35 KDa (F1, fine nanoparticles) and 280 KDa–450 nm (F2, larger nanoparticles), which mainly consisted of organic matter (OM), iron (Fe), and manganese (Mn) (oxy)hydroxides and clay minerals. Fine nanoparticles could strongly adsorb Cd and Pb under all environmental conditions. Mn and Fe (oxy)hydroxides generally formed under neutral to alkaline conditions and exhibited adsorption capabilities for Cd and Pb, respectively. Transport experiments were conducted under different pH and ionic strength conditions. At pH 3.0, soil colloids had little effect on the transport of Cd2+ and Pb2+. At pH 5.0, soil colloids inhibited the transport of Cd2+ by 16.1%, and Pb2+ recovery was still 0.0%. At pH 7.0 and 9.0, soil colloids facilitated the transport of Cd2+ by 15.6% and 29.6%, facilitated Pb2+ by 1.3% and 6.4%. At an ionic strength of 0, 0.005, and 0.01 mol L−1 NaNO3, soil colloids facilitated the transport of Cd2+ by 77.7%, 45.8%, and 15.6%, only facilitated the transport of Pb2+ by 46.2% at an ionic strength of 0 mol L−1 NaNO3. At an ionic strength of 0.05 mol L−1 NaNO3, soil colloids inhibited the transport of Cd2+ and Pb2+ by 33.1% and 21.0%, respectively. The transport of Cd2+ and Pb2+ facilitated by soil colloids was clearly observed under low ionic strength and non-acidic conditions, which can generate a potential environmental risk.