Field-flow fractionation is coming of age as a family of analytical methods for separating and characterizing macromolecules, nanoparticles, and particulates. The capabilities and versatility of these techniques are discussed in light of the challenges that are being addressed in analyzing nanometer-sized sample components and the insights gained through their use in applications ranging from materials science to biology. (To listen to a podcast about this feature, please go to the Analytical Chemistry multimedia page at pubs.acs.org/page/ancham/audio/index.html .).
Self-assembled cationic lipid-DNA complexes have shown an ability to facilitate the delivery of heterologous DNA across outer cell membranes and nuclear membranes (transfection) for gene therapy applications. While the size of the complex and the surface charge (which is a function of the lipid-to-DNA mass ratio) are important factors that determine transfection efficiency, lipid-DNA complex preparations are heterogeneous with respect to particle size and net charge. This heterogeneity contributes to the low transfection efficiency and instability of cationic lipid-DNA vectors. Efforts to define structure-activity relations and stable vector populations have been hampered by the lack of analytical techniques that can separate this type of particle and analyze both the physical characteristics and biological activity of the resulting fractions. In this study, we investigated the feasibility of flow field-flow fractionation (flow FFF) to separate cationic lipid-DNA complexes prepared at various lipid-DNA ratios. The compatibility of the lipid-DNA particles with several combinations of FFF carrier liquids and channel membranes was assessed. In addition, changes in elution profiles (or size distributions) were monitored as a function of time using on-line ultraviolet, multiangle light scattering, and refractive index detectors. Multiangle light scattering detected the formation of particle aggregates during storage, which were not observed with the other detectors. In comparison to population-averaged techniques, such as photon correlation spectroscopy, flow FFF allows a detailed examination of subtle changes in the physical properties of nonviral vectors and provides a basis for the definition of structure-activity relations for this novel class of pharmaceutical agents.
Thermal field-flow fractionation
(ThFFF) was designed to investigate the retention behavior of a series
of dendritic
polyethylenes synthesized using a chain walking catalyst (cwPE) with
variations in the branching architecture. The retention behavior of
these macromolecules correlates with their branching. Based on differences
in the Soret coefficient, a new model has been developed for the application
of ThFFF as an alternative to the branching calculation approach based
on light scattering or viscosity for the branching analysis of novel
short-chain branched PEs.
The purpose of this study is to develop a novel bacterial analysis method by coupling the flow field-flow fractionation (flow FFF) separation technique with detection by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. The composition of carrier liquid used for flow FFF was selected based on retention of bacterial cells and compatibility with the MALDI process. The coupling of flow FFF and MALDI-TOF MS was demonstrated for P. putida and E. coli. Fractions of the whole cells were collected after separation by FFF and further analyzed by MALDI-MS. Each fraction, collected over different time intervals, corresponded to different sizes and possibly different growth stages of bacteria. The bacterial analysis by flow FFF/MALDI-TOF MS was completed within 1 h with only preliminary optimization of the process.
Decomposition of a fuel-soluble precursor was used for in situ generation of Pd/PdO nanoparticles, which then catalyzed ignition of the methane/O2/N2 flow. To help understand the relationship between particle properties and activity, the composition, structure, and surface chemical state of the particles were determined by a combination of high-resolution transmission electron microscopy (HRTEM), electron diffraction, scanning transmission electron microscopy/energy dispersive X-ray spectroscopy (STEM/EDX), and X-ray photoelectron spectroscopy (XPS). The particles, collected under methane-free conditions, were found to be primarily crystalline, metallic Pd, with TEM results showing a narrow size distribution around 8 nm and scanning mobility particle sizing measurements (SMPS) indicating a median particle size of ∼10 nm. The ignition temperature was lowered ∼150 K by the catalyst, and we present evidence that ignition is correlated with formation of a subnanometer oxidized Pd surface layer.
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