The accurate analysis of silver nanoparticles (AgNPs) in complicated environmental and biological matrix is the premise for the assessment of AgNP risks. In the past few decades, a myriad of methods have been developed for the concentration and determination of AgNPs. In this chapter, methodologies for the separation, characterization, and quantification of AgNPs are introduced, and the advantages and shortcomings of each technique are also discussed. In most cases, multiple schemes are often needed to get comprehensive information of the analytes. To meet the ultra-trace detection of AgNPs in the environment, techniques with higher resolution and sensitivity are required.