Colloidal hybrid nanoparticles integrate two or more material domains into a single construct to achieve multifunctionality and synergistic properties. The synthesis of these complex multicomponent hybrid nanoparticles often yields size and composition distributions that subsequently impact observed properties and complicate structure–function studies. Analyses of nanoparticle systems such as these that contain multiple simultaneously occurring distributions are highly challenging. In this work, we introduce thermal field-flow fractionation (ThFFF) as an analytical method to separate and characterize iron oxide (Fe3O4), platinum (Pt), and multicomponent platinum–iron oxide (Pt–Fe3O4) hybrid nanoparticles by composition. The transport of matter by a temperature gradient or thermal diffusion is exploited in the ThFFF separation and characterization of inorganic nanoparticles. Thermal diffusion coefficient (DT ) values are determined from measured ThFFF retention times and translational diffusion coefficients. The DT for pure Fe3O4 and pure Pt are found to bracket those determined for hybrid Pt–Fe3O4 nanoparticles, demonstrating a proportionality between DT and composition. Furthermore, composition distributions can be determined since measured DT values vary as a function of retention time. The correlation of DT -derived cumulative distribution plots with TEM-EDS results confirmed the compositional polydispersity within hybrid nanoparticle populations. This composition-based fractionation of multicomponent inorganic nanoparticles provides insights into the fundamental forces governing thermal diffusion and highlights the ability of this technique for complex nanoparticle separations and analyses.

The impact of asymmetrical flow field-flow fractionation (AF4) on protein aggregate species is investigated with the aid of multiangle light scattering (MALS) and dynamic light scattering (DLS). The experimental parameters probed in this study include aggregate stability in different carrier liquids, shear stress (related to sample injection), sample concentration (during AF4 focusing), and sample dilution (during separation). Two anti-streptavidin (anti-SA) IgG1 samples composed of low and high molar mass (M) aggregates are subjected to different AF4 conditions. Aggregates suspended and separated in phosphate buffer are observed to dissociate almost entirely to monomer. However, aggregates in citric acid buffer are partially stable with dissociation to 25% and 5% monomer for the low and high M samples, respectively. These results demonstrate that different carrier liquids change the aggregate stability and low M aggregates can behave differently than their larger counterparts. Increasing the duration of the AF4 focusing step showed no significant changes in the percent monomer, percent aggregates, or the average Ms in either sample. Syringe-induced shear related to sample injection resulted in an increase in hydrodynamic diameter (dh) as measured by batch mode DLS. Finally, calculations showed that dilution during AF4 separation is significantly lower than in size exclusion chromatography with dilution occurring mainly at the AF4 channel outlet and not during the separation. This has important ramifications when analyzing aggregates that rapidly dissociate (<∼2s) upon dilution as the size calculated by AF4 theory may be more accurate than that measured by online DLS. Experimentally, the dhs determined by online DLS generally agreed with AF4 theory except for the more well retained larger aggregates for which DLS showed smaller sizes. These results highlight the importance of using AF4 retention theory to understand the impacts of dilution on analytes.

The design and performance of a semi-preparative asymmetrical flow field-flow fractionation (SP-AF4) channel are investigated with the objective of better understanding and exploiting the relationship between channel dimensions, sample loading, and resolution. Most size-based separations of nanometer and submicrometer particles are currently limited to analytical scale quantities (<100μg). However, there is a strong need to fractionate and collect larger quantities so that fundamental properties of the more narrowly dispersed fractions can be studied using additional characterization methods and for subsequent applications. In this work, dimensions of the spacer that defines the form of SP-AF4 channels are varied and their performances are assessed with respect to sample focusing position and loading. Separations are performed in aqueous and organic carrier fluids. A critical evaluation of channel dimensions showed that increasing the channel breadth is a practical and effective route to maintaining separation resolution while increasing sample loads to milligram quantities. Good size resolution (∼1.0) is achieved for separations of 10mg of 50 and 100nm silica nanoparticles suspended in water and up to 0.6mg of ∼10 to 35nm inorganic hybrid nanoparticles suspended in tetrahydrofuran. This work represents important advances in the understanding of SP-AF4 separations and extends sample loading capacities in both aqueous and organic solvents.

High and low-density lipoproteins (HDL and LDL) are attractive targets for biomarker discovery. However, ultracentrifugation (UC), the current methodology of choice for isolating HDL and LDL is tedious, requires large sample volume, results in sample loss, and does not readily provide information on particle size. In this work, human plasma HDL and LDL are separated and collected using semi-preparative asymmetrical flow field-flow fractionation (SPAF4) and UC. The SP-AF4 and UC separation conditions, sample throughput, and liquid chromatography/mass spectrometry (LC/MS) lipidomic results are compared. Over 600 μg of total proteins are recovered in a single SP-AF4 run and Western blot results confirm apoA1 pure and apoB100 pure fractions, consistent with HDL and LDL, respectively. The SP-AF4 separation requires ~60 minutes per sample, thus providing a marked improvement over UC which can span hours to days. Lipidome analysis of SP-AF4 prepared HDL and LDL fractions are compared to UC prepared HDL and LDL samples. Over 270 lipids in positive MS mode and over 140 lipids in negative MS mode are identified by both sample preparation techniques with over 98% overlap between the lipidome. Additionally, lipoprotein size distributions are determined using analytical scale AF4 coupled with multiangle light scattering (MALS) and dynamic light scattering (DLS) detectors. These developments position SP-AF4 as a sample preparation method of choice for lipoprotein biomarker characterization and identification.

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

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Field-flow fractionation (FFF) is a family of techniques that was created especially for separating and characterizing macromolecules, nanoparticles, and micrometer-sized analytes. It is coming of age as new nanomaterials, polymers, composites, and biohybrids with remarkable properties are introduced and new analytical challenges arise due to synthesis heterogeneities and the motivation to correlate analyte properties with observed performance. Appreciation of the complexity of biological, pharmaceutical, and food systems and the need to monitor multiple components across many size scales have also contributed to FFF's growth. This review highlights recent advances in FFF capabilities, instrumentation, and applications that feature the unique characteristics of different FFF techniques in determining a variety of information, such as averages and distributions in size, composition, shape, architecture, and microstructure and in investigating transformations and function. Expected final online publication date for the Annual Review of Analytical Chemistry, Volume 14 is June 2021. Please see http://www.annualreviews.org/page/journal/pubdates for revised estimates.

This paper reports the first use of a remote sensing device to measure emissions from in-use marine vessels. Emissions from 307 small marine vessels were measured as they passed through the Hiram M. Chittenden locks near Seattle, WA. Of these vessels, 89 were matched to state registration information to allow for further analysis of emissions vs model year, fuel type, and engine type. Emission factors are reported for CO, HC, and NOx in grams of pollutant per kilogram of fuel. The measured emission factors generally agreed with those derived from laboratory studies. HC emissions are disproportionately skewed across the fleet where 40% of the emissions come from just 10% of the fleet. These are most likely due to the remaining two-stroke engines in the fleet. CO and HC emissions show no improvement with newer vessels.