Lychnophora salicifolia, commonly known as "arnicão", is used as an anti-inflammatory agent and as a flavoring agent in the Brazilian traditional spirit "cachaça". In this work, the permeation process of vicenin-2 (1) and lychnopholic acid (2) (major secondary metabolites from the hydroalcoholic extract) was investigated using Caco-2 cells. For this investigation, a new HPLC-DAD method was developed and validated for the quantification step. It was observed that 2 crosses the Caco-2 cell monolayer by passive diffusion. On the other hand, 1 was not transported, suggesting no absorption and no efflux of this compound in Caco-2 cells.