The family Chironomidae belongs to the most abundant macroinvertebrates in samples for water quality assessment and displays a wide tolerance range to contaminants, which makes it an excellent bioindicator. The species Genus 1 sp. 2 (Chironomidae: Orthocladiinae), included among the larval keys of the Cricotopus-Oliveiriella complex, is difficult to determine based on its larval instar using the current morphological keys, which makes it necessary to use pupae for a species-level identification. In this study, 103 organisms in the IV larval instar were collected from tributaries of the high Chinchiná river basin (Caldas-Colombia), along with eight organisms at the pupal level (reared in the laboratory).