Updated project metadata. Trypanosoma cruzi metacyclogenesis is a natural process that occurs inside triatomine insect vector and corresponds to the differentiation of non-infective epimastigotes into infective metacyclic trypomastigotes. Here, centered on LC-MS/MS approach, a large scale phospho (proteome) study was carried out aiming to quantitatively understand the metacyclogenesis. Cells were colected along metacyclogenesis totalizing six points, stress 2 hours, adhered 12, 24 and 48 hours, and metacyclic trypomastigote from two biological replicates. After extraction, total protein were digested, fractioned and analyzed by LC-MS/MS. The samples for phosphoproteomics were processed for phosphopeptide enrichment prior to the analysis.