Summary Setenta and five non-metric coronal features in permanent and deciduous dentition were registered in 178 dental models of Afro-Colombian individuals belonging to the Guapi community. Comparative data were collected from 33 African groups, another Afro-American group in permanent dentition, from 4 Afro-American groups and 12 other groups of different ethnic descent in deciduous dentition. A multivariate analysis of major components based on dental frequencies showed that Afro-Colombian people have great genetic affinity with Africa Sub-Sahariana, specifically with groups from the West, the East and the South East. The same analysis identified a marked dental divergence with North African populations. The results of this research are in line with recent genetic studies, based on ADNmt markers. Although dentition is evolutionary conservative and stable on a short diachronistic scale, it was concluded that in 400 years of microevolution of Africans in Colombia, their original genetic pool has been slightly modified, due to the influence of selective pressures and inter-ethnic gene flow, with American natives and groups of captive origin. It is suggested that Afro-descendant groups from North, Central and South America have different population and evolving histories, due both to regional and local phenotypic differentiation processes and to different geographical and ethnic origins. Abstract Seventy-five Crown non-metric traits in the permanent and decisive dentition were drawn in 178 dental Casts of Afro-Colombians from Guapi. Comparative data from 33 African groups, another African-American group in the permanent dentition, 4 African-American populations and other 12 human populations of different ancestry in the decisive dentition had gathered. A multivariate analysis of main components (PCA) based on dental trait frequencies, managed that Afro-Colombians have close genetic concerns with Sub-Saharan African populations, specifically with groups from Western, Eastern and South-Eastern Africa. This analysis also detected a strong dental divergence between North African groups and Afro-Colombians. These results agreed with recent mtDNA studies. The human dentition is evolving conservative and stable, however, in 400 years of Africans evolution in Colombia. Select pressures and inter-ethnic gene flow with Amerindian and Caucasian populations slowly modified the original African gene pool of the Afro-Colombian community. Finally, this study studies that African-Americans from North, Caribbean-Central and South America have different evolutionary and populational histories due to local and regional processes of phenotypic differentiation, as well as to different geography and ethnic origins.