Analytical approaches for characterizing and quantifying engineered nanoparticles in biological matrices from an (eco)toxicological perspective: old challenges, new methods and techniques

: To promote the safer by design strategy and assess environmental risks of engineered nanoparticles (ENPs), it is essential to understand the fate of ENPs within organisms. This understanding in living organisms is limited by challenges in characterizing and quantifying ENPs in biological media. Relevant literature in this area is scattered across research from the past decade or so, and it consists mostly of medically oriented studies. This review first introduces those modern techniques and methods that can be used to extract, characterize, and quantify ENPs in biological matrices for (eco)toxicological purposes. It then summarizes recent research developments within those areas most relevant to the context and field that are the subject of this review paper. These comprise numerous in-situ techniques and some ex-situ techniques. The former group includes techniques allowing to observe specimens in their natural hydrated state (e.g., scanning electron microscopy working in cryo mode and high-pressure freezing) and microscopy equipped with elemental microanalysis (e.g., energy-dispersive X-ray spectroscopy); two-photon laser and coherent anti-Stokes Raman scattering microscopy; absorption-edge synchrotron X-ray computed microtomography; and laser ablation-inductively coupled plasma mass spectrometry (LA-ICP-MS). The latter group includes asymmetric flow field flow fractionation coupled with ICP-MS and single particle-ICP-MS. Our review found that most of the evidence gathered for ENPs actually focused on a few metal-based ENPs and carbon nanotube and points to total mass concentration but no other particles properties, such as size and number. Based on the obtained knowledge, we developed and presented a decision scheme and analytical toolbox to help orient scientists toward selecting appropriate ways for investigating the (eco)toxicity of ENPs that are consistent with their properties