The biodiversity of the tropical Americas is a consequence of the interplay between geological and climatic events, with the Andean uplift being a major driver of speciation. Multiple studies have shown that species diversification promoted by the Andes can occur in the presence or absence of gene flow. However, to date, the majority of research addressing this aspect has been conducted in vertebrates, whereas other highly diverse tropical organisms such as arthropods remain uninvestigated. We used a combination of phylogenetics, population genetic analyses and species distribution models to explore whether the northern Andes played a role in the diversification of Ancylometes bogotensis. We detected two major lineages that are separated by the Eastern Cordillera of the Colombian Andes, and they share the same climatic niche. These groups diverged at ~3.85 Mya and exhibit no signatures of gene flow, which can be a consequence of the Andean highlands being poorly suited habitats for this species, thus preventing their genetic connectivity. Our study reveals that the genetic structure of an arachnid species that has limited dispersal capacity and is highly dependent on water bodies is shaped by the Andean orogeny. The generality of this observation remains to be assessed in other invertebrates.