Protocols for sample preparation and measurement of nanoparticle stability and their biotransformation in biological matrices.

This report provides protocols for sample preparation and measurement of nanoparticle stability and biotransformation in biological matrices developed in the scope of the MetrINo project. The protocols focus on optimised matrix spiking, including minimisation of matrix interferences, and extraction of nanomaterials from model biological matrices. This report describes the optimised spiking protocols for selected NPs (nanoparticles, both liposomes and inorganic NPs) into biological matrices of increasing complexity (from simple PBS buffer to highly complex biological serum), as well as NP extraction from these matrices. In addition to chemical and enzymatic protocols, fractionation protocols have been developed to separate NPs from the protein fractions within the complex matrices using asymmetrical-flow field-flow fractionation (AF4) methods, which provide sufficiently high material recovery rates and negate the need for more invasive alternative extraction methods. Example data sets obtained using the optimised protocols are also provided.