The frequent presence of smoking pipes in Early Ceramic Period archaeological sites in central Chile and the large body of evidence on the smoking of Nicotiana spp. in prehispanic sites elsewhere in the Americas call for a methodology to identify Nicotiana species used in Chile. We have performed a morphometric study on seeds of Nicotiana species which serves as a reference against which archaeological seeds may be compared. Seed shape and size, ornamentation pattern of epidermal cells, and location of the hilum were relevant parameters for reliable taxonomic determinations. Using these characters, we identified Nicotiana seeds found in Las Morrenas 1 archaeological site in central Chile as N. corymbosa.