Quantitative size-resolved characterization of mRNA nanoparticles by in-line coupling of asymmetrical-flow field-flow fractionation with small angle X-ray scattering.

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.

Novel Benchtop Magnetic Particle Spectrometer for Process Monitoring of Magnetic Nanoparticle Synthesis.

Magnetic nanoparticles combine unique magnetic properties that can be used in a variety of biomedical applications for therapy and diagnostics. These applications place high demands on the magnetic properties of nanoparticles. Thus, research, development, and quality assurance of magnetic nanoparticles requires powerful analytical methods that are capable of detecting relevant structural and, above all, magnetic parameters. By directly coupling nanoparticle synthesis with magnetic detectors, relevant nanoparticle properties can be obtained and evaluated, and adjustments can be made to the manufacturing process in real time. This work presents a sensitive and fast magnetic detector for online characterization of magnetic nanoparticles during their continuous micromixer synthesis. The detector is based on the measurement of the nonlinear dynamic magnetic response of magnetic nanoparticles exposed to an oscillating excitation at a frequency of 25 kHz, a technique also known as magnetic particle spectroscopy. Our results underline the excellent suitability of the developed magnetic online detection for coupling with magnetic nanoparticle synthesis based on the micromixer approach. The proven practicability and reliability of the detector for process monitoring forms the basis for further application fields, e.g., as a monitoring tool for chromatographic separation processes.

Asymmetrical Flow Field-Flow Fractionation for Sizing of Gold Nanoparticles in Suspension.

Particle size is arguably the most important physico-chemical parameter associated with the notion of a nanoparticle. Precise knowledge of the size and size distribution of nanoparticles is of utmost importance for various applications. The size range is also important, as it defines the most "active" component of a nanoparticle dose. Asymmetrical Flow Field-Flow Fractionation (AF4) is a powerful technique for sizing of particles in suspension in the size range of approximately 1-1000 nm. There are several ways to derive size information from an AF4 experiment. Besides coupling AF4 online with size-sensitive detectors based on the principles of Multi-Angle Light Scattering or Dynamic Light Scattering, there is also the possibility to correlate the size of a sample with its retention time using a well-established theoretical approach (FFF theory) or by comparing it with the retention times of well-defined particle size standards (external size calibration). We here describe the development and in-house validation of a standard operating procedure (SOP) for sizing of an unknown gold nanoparticle sample by AF4 coupled with UV-vis detection using external size calibration with gold nanoparticle standards in the size range of 20-100 nm. This procedure provides a detailed description of the developed workflow including sample preparation, AF4 instrument setup and qualification, AF4 method development and fractionation of the unknown gold nanoparticle sample, as well as the correlation of the obtained results with the established external size calibration. The SOP described here was eventually successfully validated in the frame of an interlaboratory comparison study highlighting the excellent robustness and reliability of AF4 for sizing of nanoparticulate samples in suspension.

Nanoplastic Analysis by Online Coupling of Raman Microscopy and Field-Flow Fractionation Enabled by Optical Tweezers.

Nanoplastic pollution is an emerging environmental concern, but current analytical approaches are facing limitations in this size range. However, the coupling of nanoparticle separation with chemical characterization bears potential to close this gap. Here, we realize the hyphenation of particle separation/characterization (field-flow fractionation (FFF), UV, and multiangle light scattering) with subsequent chemical identification by online Raman microspectroscopy (RM). The problem of low Raman scattering was overcome by trapping particles with 2D optical tweezers. This setup enabled RM to identify particles of different materials (polymers and inorganic) in the size range from 200 nm to 5 µm, with concentrations in the order of 1 mg/L (109 particles L-1). The hyphenation was realized for asymmetric flow FFF and centrifugal FFF, which separate particles on the basis of different properties. This technique shows potential for application in nanoplastic analysis, as well as many other fields of nanomaterial characterization.

Thermal Decoherence of a Nonequilibrium Polariton Fluid.

Exciton polaritons constitute a unique realization of a quantum fluid interacting with its environment. Using selenide-based microcavities, we exploit this feature to warm up a polariton condensate in a controlled way and monitor its spatial coherence. We determine directly the amount of heat picked up by the condensate by measuring the phonon-polariton scattering rate and comparing it with the loss rate. We find that, upon increasing the heating rate, the spatial coherence length decreases markedly, while localized phase structures vanish, in good agreement with a stochastic mean-field theory. From the thermodynamical point of view, this regime is unique, as it involves a nonequilibrium quantum fluid with no well-defined temperature but which is nevertheless able to pick up heat with dramatic effects on the order parameter.

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

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

Tunable Bragg polaritons and nonlinear emission from a hybrid metal-unfolded ZnSe-based microcavity.

Strong light-matter interaction in Bragg structures possesses several advantages over conventional microcavity system. These structures provide an opportunity to incorporate a large number of quantum wells without increasing the mode volume. Further, it is expected that the strong coupling could occur over the entire thickness of the Bragg structure, and the system offers an improved overlap between exciton wave function and light mode. However, advanced experiments in Bragg structures require a precise control and manipulation of quantum states of Bragg polaritons. Here, we propose and experimentally demonstrate novel methods for the modulation of Bragg polariton eigenstates. The modulation will be shown to even exceed 10 meV if the thickness of the top layer of the ZnSe-based Bragg structure is changed or if a thin silver layer is deposited on top of the structure. The Q value of the Bragg mode will be enhanced by a factor of 2.3 for a 30 nm silver layer. In addition, we report on the observation of nonlinear emission of the lower Bragg polariton mode in the hybrid structure being achieved when excitation dependent measurements are performed. Our results open the door to create a confined Bragg polariton system similar to conventional microcavities.

Observation of a hybrid state of Tamm plasmons and microcavity exciton polaritons.

We present evidence for the existence of a hybrid state of Tamm plasmons and microcavity exciton polaritons in a II-VI material based microcavity sample covered with an Ag metal layer. The bare cavity mode shows a characteristic anticrossing with the Tamm-plasmon mode, when microreflectivity measurements are performed for different detunings between the Tamm plasmon and the cavity mode. When the Tamm-plasmon mode is in resonance with the cavity polariton four hybrid eigenstates are observed due to the coupling of the cavity-photon mode, the Tamm-plasmon mode, and the heavy- and light-hole excitons. If the bare Tamm-plasmon mode is tuned, these resonances will exhibit three anticrossings. Experimental results are in good agreement with calculations based on the transfer matrix method as well as on the coupled-oscillators model. The lowest hybrid eigenstate is observed to be red shifted by about 13 meV with respect to the lower cavity polariton state when the Tamm plasmon is resonantly coupled with the cavity polariton. This spectral shift which is caused by the metal layer can be used to create a trapping potential channel for the polaritons. Such channels can guide the polariton propagation similar to one-dimensional polariton wires.

Advanced analysis of polymer emulsions: Particle size and particle size distribution by field-flow fractionation and dynamic light scattering.

Field flow fractionation (FFF) is an advanced fractionation technique for the analyses of very sensitive particles. In this study, different FFF techniques were used for the fractionation and analysis of polymer emulsions/latexes. As model systems, a pure acrylic emulsion and emulsions containing titanium dioxide were prepared and analyzed. An acrylic emulsion polymerization was conducted, continuously sampled from the reactor and subsequently analyzed to determine the particle size, radius of gyration in specific, of the latex particles throughout the polymerization reaction. Asymmetrical flow field-flow fractionation (AF4) and sedimentation field-flow fractionation (SdFFF), coupled to a multidetector system, multi-angle laser light scattering (MALLS), ultraviolet (UV) and refractive index (RI), respectively, were used to investigate the evolution of particle sizes and particle size distributions (PSDs) as the polymerization progressed. The obtained particle sizes were compared against batch-mode dynamic light scattering (DLS). Results indicated differences between AF4 and DLS results due to DLS taking hydration layers into account, whereas both AF4 and SdFFF were coupled to MALLS detection, hence not taking the hydration layer into account for size determination. SdFFF has additional separation capabilities with a much higher resolution compared to AF4. The calculated radii values were 5 nm larger for SdFFF measurements for each analyzed sample against the corresponding AF4 values. Additionally a low particle size shoulder was observed for SdFFF indicating bimodality in the reactor very early during the polymerization reaction. Furthermore, different emulsions were mixed with inorganic species used as additives in cosmetics and coatings such as TiO2. These complex mixtures of species were analyzed to investigate the retention and particle interaction behavior under different AF4 experimental conditions, such as the mobile phase. The AF4 system was coupled online to inductively coupled plasma mass spectrometry (ICP-MS) for elemental speciation and identification of the inorganic additive. SdFFF had a larger separation power to distinguish different particle size populations whereas AF4 had the capability of separating the organic particles and inorganic TiO2 particles, with high resolution.

Exciton-polariton gas as a nonequilibrium coolant.

Using angle-resolved Raman spectroscopy, we show that a resonantly excited ground-state exciton-polariton fluid behaves like a nonequilibrium coolant for its host solid-state semiconductor microcavity. With this optical technique, we obtain a detailed measurement of the thermal fluxes generated by the pumped polaritons. We thus find a maximum cooling power for a cryostat temperature of 50 K and below where optical cooling is usually suppressed, and we identify the participation of an ultrafast cooling mechanism. We also show that the nonequilibrium character of polaritons constitutes an unexpected resource: each scattering event can remove more heat from the solid than would be normally allowed using a thermal fluid with normal internal equilibration.

Colloid and heavy metal transport at landfill sites in direct contact with groundwater.

Colloids are ubiquitous in aquatic systems and are suspected of facilitating contaminant transport. At sites where the disposed waste is in direct contact with ground water, two main prerequisites for colloidal transport are fulfilled: these two prerequisites are a high concentration of colloids and many different contaminants, some that are very unlikely to be transported in an aqueous solution. In our investigation, three landfill sites with different historical background and hydrogeological conditions were examined. The colloids upstream, downstream, and inside the waste disposal sites were characterized with respect to their size distribution and chemical composition. The particle concentration upgradient and downgradient of the sites were 2-23 and 8-80 mg/L, respectively. Inside the waste disposal sites the particle concentration was 160-870 mg/L. The particles upgradient reflected the mineral composition of the aquifer, with calcareous colloids and silicates dominating the composition. Downgradient of the sites, we saw an increase of iron-precipitates and salt colloids, together with organic colloids. However, the downgradient colloids were significantly different from the colloids and particles inside of the disposal sites with respect to their size and chemical composition. Colloids inside the disposal sites reflected the waste composition and degradation. The association of heavy metal ions to colloids and particles showed a surprisingly high fraction of dissolved metal ions. We determined that the lowest metal ion concentrations (less than 20% of the total concentration) were associated with the colloid size class between 10 nm and 1 microm, which is considered most mobile in porous systems. The association of Fe and Mn to colloids was dominated by the redox conditions inside the disposal sites, where there was a reducing environment, Fe and Mn were dissolved. Outside the disposal sites, where there was an oxidizing environment, these metals formed colloids and particles >1 microm. Together with these particles, As was precipitating. For other metal ions (Cd, Co, Cu, Ni, Pb, Zn), we determined an association to colloids coinciding roughly with the colloid size distribution. The results suggest that the change of hydrochemical conditions at the interface, from a reducing, high ionic strength environment inside of the disposal sites to an oxidizing, low ionic strength environment in the groundwater together with physical filtration effects for the larger particles, is an effective chemical barrier for colloids. Field observations suggest, that the colloids form a rather persistent coating around the aquifer matrix that reduces the hydraulic conductivity and enhances the sorption capacity of the aquifer close to the waste disposal sites. In every case, there was an increase of the contaminant concentrations downstream of the waste disposal sites, but the increase was less than expected from the initial transport calculations, which were under the assumption that there was a direct contact between the waste disposal site and the groundwater. It seems more than likely, that under the given conditions, colloids are contributing to a self sealing layer at the landfill bottom. The results of this study are relevant for the assessment of many landfills without appropriate landfill sealing systems.