NBL was established by the Atomic Energy Commission in 1949 in New Brunswick, NJ. It was initially staffed by scientists from the National Bureau of Standards that had contributed to the measurement science of nuclear materials for the Manhattan Project. NBL's initial mission was to provide a Federal capability for the assay of uranium-containing materials for the nation's developing atomic energy program. Over the years NBL expanded its capabilities, developing newer and improved methods and procedures, and certifying additional reference materials for use around the world. The capability for plutonium measurements was implemented at NBL in 1959. NBL was relocated from New Jersey to the site at Argonne National Laboratory during the period 1975-77.

Since its beginning, NBL has maintained a Center of Excellence in the analytical chemistry and measurement science of nuclear materials. In this role, NBL continues to perform state-of-the-art measurements of the elemental and isotopic compositions for a wide range of nuclear materials.