Method for Extraction and Quantification of Metal-Based Nanoparticles in Biological Media: Number-Based...

A multistep sample preparation method was developed to separate metal-based engineered nanoparticles (ENPs) from biological samples. The method was developed using spiked zebrafish tissues and standard titanium dioxide (TiO₂) and cerium dioxide (CeO₂) ENPs. Single-particle inductively coupled plasma mass spectrometry was used to quantify the separated particles in terms of number concentration. This method demonstrated mass recoveries of more than 90% and did not strikingly alter the median particles size. High number recoveries were calculated for CeO₂ ENPs (>84%). Particle number recoveries were poor for TiO₂ ENPs (<25%), which could be due to the interference of ⁴⁸Ca with the measured isotope ⁴⁸Ti. The method was verified using zebrafish exposed to CeO₂ ENPs to test its applicability for nanotoxicokinetic investigations. Total mass of Ce and particle number concentration of CeO₂ ENPs were measured in different tissues. Notably, the mass-based biodistribution of Ce in the tissues did not follow the number-based biodistribution of CeO₂. Moreover, the calculated mass-based bioconcentration factors showed a different pattern in comparison to the number-based bioconcentration factors. Our findings suggest that considering mass as the sole dose-metric may not provide sufficient information to investigate toxicity and toxicokinetics of ENPs.