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Objective was to develop and characterize nano-edetate calcium disodium (Ca-Na2EDTA) dry powder inhaler (DPI), and assess its in vitro and in vivo deposition using pharmacoscintigraphy techniques. Factors influencing nanoparticle formation including concentration of drug, polymer solution and stirring rate were determined. Optimized formulation was characterized with the help of SEM, TEM and Malvern Zetasizer studies. Any change in physical characteristics after nanosizing was determined by FT-IR, XRD and DSC studies. Anderson cascade impaction showed that nano Ca-Na2EDTA exhibited significantly higher respirable fraction of 67.35 ± 2.27% and 66.40 ± 2.87% by scintigraphic and spectroscopic analysis respectively, as compared to 10.08 ± 1.17% and 9.36 ± 1.02% respectively for micronized form. Ventilation lung scintigraphy done in 12 volunteers showed significant increase in drug delivery till alveolar region with nano Ca-Na2EDTA. The developed formulation may have a role in neutralizing heavy metal toxicity through inhalation route, including radio-metal contamination.