Validity range of centrifuges for the regulation of nanomaterials: from classification to as-tested coronas

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Abstract
Granulometry is the regulatory category where the differences between traditional materials and nanomaterials culminate. Reported herein is a careful validation of methods for the quantification of dispersability and size distribution in relevant media, and for the classification according to the EC nanodefinition recommendation. Suspension-based techniques can assess the nanodefinition only if the material in question is reasonably well dispersed. Using dispersed material of several chemical compositions (organic, metal, metal-oxide) as test cases we benchmark analytical ultracentrifugation (AUC), dynamic light scattering (DLS), hydrodynamic chromatography, nanoparticle tracking analysis (NTA) against the known content of bimodal suspensions in the commercially relevant range between 20 nm and a few microns. The results validate fractionating techniques, especially AUC, which successfully identifies any dispersed nanoparticle content from 14 to 99.9 nb% with less than 5 nb% deviation. In contrast, our screening casts severe doubt over the reliability of ensemble (scattering) techniques and highlights the potential of NTA to develop into a counting upgrade of DLS. The unique asset of centrifuges with interference, X-ray or absorption detectors—to quantify the dispersed solid content for each size interval from proteins over individualized nanoparticles up to agglomerates, while accounting for their loose packing—addresses also the adsorption/depletion of proteins and (de-)agglomeration of nanomaterials under cell culture conditions as tested for toxicological endpoints.

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