Understanding the Significance of Sample Preparation in Studies of the Nanoparticle Metabolite Corona

[Image: see text] The adsorption of metabolites to the surface of nanomaterials is a growing area of interest in the field of bionanointeractions. Like its more-established protein counterpart, it is thought that the metabolite corona has a key role in the uptake, distribution, and toxicity of nanomaterials in organisms. Previous research has demonstrated that nanomaterials obtain a unique metabolite fingerprint when exposed to biological matrices; however, there have been some concerns raised over the reproducibility of bionanointeraction research due to challenges in dispersion of nanomaterials and their stability. As such, this work investigates a much-overlooked aspect of this field, i.e., sample preparation, which is vital to the accurate, reproducible, and informative analysis of the metabolite corona. The impact of elution buffer pH, volume, and ionic strength on the metabolite corona composition acquired by uncapped and polyvinylpyrrolidone (PVP)-capped TiO(2) from mixtures of cationic and anionic metabolites was studied. We demonstrate the temporal evolution of the TiO(2) metabolite corona and the recovery of the metabolite corona, which resulted from a complex biological matrix, in this case human plasma. This work also demonstrates that it is vital to optimize sample preparation for each nanomaterial being investigated, as the metabolite recovery from Fe(3)O(4) and Dispex-capped TiO(2) nanomaterials is significantly reduced compared to the aforementioned uncapped and PVP-capped TiO(2) nanomaterials. These are important findings for future bionanointeraction studies, which is a rapidly emerging area of research in nanoscience.