In this study, a method for separation, size characterization, and speciation of gold and silver nanoparticles was developed through the use of micellar electrokinetic chromatography (MEKC) coupled to inductively coupled plasma-mass spectrometry (ICP-MS) for the first time. Figures of merit in this proof-of-principle study include peak area precision of 4-6%, stable migration times (1.4% with internal standard), and capillary recoveries on the order of 72-100% depending on species and nanoparticle size, respectively. Detection limits are currently in the sub-microgram per liter range. For example, a total of 1500 50-nm-sized gold nanoparticles were successfully detected. After careful optimization, MEKC-ICP-MS was used to separate engineered nanoparticles (ENPs) of different composition. Speciation analysis of ENPs and free metal ions in solution was feasible using a complexing agent (penicillamine). Gold speciation analysis of a dietary supplement, which contained approximately 6-nm-sized gold nanoparticles, was demonstrated.