Chromatography is emerging as an efficient technique for the separation of engineered nanoparticles (NPs) and has been gaining tremendous attention due to its ease, facile, and cost-effective nature. This chapter puts together the various chromatography techniques so far implemented to separate or fractionate various NPs in detail. The influence of morphological features of the NPs along with overall surface properties that can aid their separation using chromatography will be highlighted. Finally, this chapter also provides an overview of the various analytical and advanced physical characterization techniques often used to evaluate the successful separation of nanoparticles.