Recently, the scope, regulation, legislation, and metrology of the analytical chemistry of engineered nanomaterials (ENMs) have been reviewed in the Part 1 of the IUPAC Technical Report. Chemical analysis of nanomaterials in complex sample matrices presents a substantial challenge for analytical science and regulatory agencies. The purpose of the present Part 2 is to discuss the detection, characterization, and quantification of nanomaterials in samples of complex matrices including methods for sample preparation and fitness for purpose. Analytical methods applied to analysis in matrices of environmental samples, food, cosmetics, and biological samples as well as those used to monitor the fate of ENMs in the environment and biological systems are reported. Tables of numerous recently published works on analyses of typical ENMs with detailed protocols and conclusive comments are presented. There is a rapid development in the field mostly in the stage of accumulation of factual material. The single-particle inductively coupled plasma mass spectrometry is already widely used at the chemical analysis of metal-containing nanoparticles.