Dual (oxygen and nitrogen) isotopic composition of the museum archived nitrates from the United States of America, South Africa and Australia was studied. The analyzed specimens were collected in middle 19th to early 20th centuries, and represent world-wide acquisition of the Smithsonian Institution Natural Museum of Natural History (Washington, D. C., USA) and the Natural History Museum (London, UK). The samples consist of transparent to semi-transparent aggregates of minute nitrate, euhedral crystallites which imply precipitation from percolating fluids under ample space and dry regimes. The major nitrate chemistry is saltpetre (KNO) with minor nitratine (NaNO). A binary plot of δN vs. δO of almost all nitrates indicates a trend, reflecting microbial origin through nitrification of ammonium. The diagram excludes the contribution of meteoric origin formed by mass-independent, photochemical reaction of NO with ozone in stratosphere. Calculated paleo-ambient fluid compositions responsible for microbial nitrification imply extreme evaporative concentration of relevant fluids under dry climatic regimes in the Northern Cape Province (South Africa) and in the Northern Territory (central Australia), and even throughout the United States of America. The dual isotopic characterization provides direct evidence to the origin of the museum archived nitrates.