Characterizing a spheroidal nanocage drug delivery vesicle using multi-detector hydrodynamic chromatography

The biological application of nanoparticles has resulted in an increased need for the development of robust, accurate, and precise methods for quality control analysis and characterization. Parameters such as particle size, particle shape, and their distributions affect end-use properties such as chemical reactivity, diffusivity, permeability, and transport. Introduced here is a hydrodynamic chromatography (HDC) method utilizing multi-angle static light scattering, quasi-elastic light scattering, differential viscometry, and differential refractometry detection for characterizing nanoscale vesicles. Quadruple-detector HDC was used to determine multiple sizing parameters and their statistical moments and distributions. Molar mass and molar mass averages were determined in a calibrant-independent fashion. Both the sizing parameters and the molar mass were measured across the HDC elution profile. The shape and structure of the nanoparticle were monitored as a function of HDC elution volume through the dimensionless ratio ρ≡R(G,z)/R(H,z). The HDC results were comparable to those obtained by transmission electron microscopy, but more extensive characterization was possible by HDC, which provided distributions of both particle size and particle shape.