This paper aims to analyse the spatial patterns of sampling effort and species richness of pteridophyte in a well-investigated region as Tuscany, Italy, by using data stored from a geodatabase storing information on the specimens preserved in the main herbaria of the region. A total of 6,905 records about pteridophyte specimens were extracted from the geodatabase, and 5,638 of such specimens were studied through the use of spatial statistical techniques. The data about the sampling effort and species richness were analysed in relation to topographical variables to assess any significant relationship. Specimen-based rarefaction techniques were used to compare areas with different number of detected species. The analysis of the sampling effort data showed a nonhomogeneous distribution of herbarium data, with some areas being intensively sampled and others being almost unsampled. Thus, the geographical distribution of specimens was extremely clustered. The comparison across geographical areas through specimen-based rarefaction curves showed great differences in species richness and sampling completeness. The analysis of the residuals of species–area relationships evidenced that the distance to water bodies was the only significant topographical variable in controlling species diversity.