Iron deficiency, when severe, leads to iron deficiency anemia (IDA) and IDA is a major health problem globally. Adding ferric pyrophosphate in the form of nanoparticles to food has the advantage of less organoleptic changes and higher bioavailability. Ferric pyrophosphate nanoparticle was synthesized and characterized as a potential candidate for food fortificant in IDA. The bioavailability of the compound has been tested in rat model along with gold standard ferrous sulphate. The LD50 of ferric pyrophosphate nanoparticle has been determined by Acute Oral Toxicity study and chronic toxicity has been tested by Repeated Oral Toxicity study. Histopathological study of organs for all animals has been done to assess iron-induced toxicity. The biomarker study for iron overdose has been done with mass spectrometry based proteomics approach.