Table 5.

Nanospecific characteristics and physico-chemical properties of drug-loaded nanoparticles, which are considered relevant for the preclinical characterization of nanomedicines/nanotherapeutics formulated in aqueous media.

Nanospecific Characteristics	Test Method
Size/size distribution	DLS
Physical form/shape/morphology	TEM, cryo-TEM
Surface charge	Zeta potential, electrophoretic mobility (EPM)
Aggregation behaviour	DLS
Stability and uniformity	DLS, UV–VIS spectroscopy
Density/weight/volume fraction of nanomaterial dispersed in the medium	Ultracentrifugation, densitometry
Drug encapsulation	UV–VIS spectrometry, high-performance liquid chromatography (HPLC)
Presence of targeting moieties	Kinetic turbidity assays, Spectroscopic assays (UV–VIS, circular dichroism), surface plasmon resonance (SPR) binding assays
Toxicity	Cytotoxicity assessment using MTT and LDH assays
Biocompatibility	Immunological response, hemolytic properties
Structural and functional properties	TEM, SEM, small-angle X-ray scattering (SAXS), NTA, high-resolution transmission electron microscopy (HRTEM), atomic force microscopy (AFM), extended X-ray absorption fine structure (EXAFS), ferromagnetic resonance (FMR), DSC, differential centrifugal sedimentation (DCS), inductively coupled plasma atomic emission spectroscopy (ICP-MS), UV–VIS, matrix-assisted laser desorption/ionization (MALDI), nuclear magnetic resonance (NMR), superparamagnetic relaxometry, tunable resistive pulse sensing (TRPS)