In order to improve our understanding of the NW-Amazonian Craton evolution, we present new petrographic, geochemical and geochronological analyses of 27 samples from the geotectonic Rio Negro-Juruena Province in eastern Colombia (Guainía and Vaupés departments). New LA-ICP-MS zircon U-Pb ages suggest that the oldest known rocks in Colombia are metamorphic rocks (migmatitic gneisses) with ages between ~ 1850 and ~ 1800 Ma, and gneisses and granitoids with ages between ~ 1800–1720 Ma which form part of the Mitú Complex, interpreted as the result of Statherian collisional and orogenic events (Querari Orogeny). Detrital zircons in low-grade meta-sedimentary sequences of the Tunuí Group (sandstones, conglomeratic sandstones and mudstones), that crop out over almost the entire basement, indicate older than ~ 1770 Ma source rocks. Intrusions of different suites of granitic rocks with syn- to post-collisional affinities suggest a termination of the collisional events between ~ 1600–1500 Ma which had affected the whole region, occasionally metasomatically overprinting parts of the Tunuí meta-sedimentary sequence. The recognizable metamorphic and magmatic-processes finish with ~ 1400–1340 Ma anorogenic granites without signs of tectonic deformation, resembling anorogenic granites in the Western Amazonian Craton in Brazil and the Parguaza Batholith in Venezuela. This study allows us to conclude that the basement records the collision of a continental arc (Rio Negro-Juruena Province) against the NW-Amazonian Craton (Ventuari-Tapajos Province) and its subsequent transition to anorogenic conditions in a continental rift setting long before the actual stable craton conditions.