As the identity of a large number of Khoe-San skeletal material in European collections recently came into question during its documentation, a re-evaluation of the remains by employing a non-invasive method such as craniometrics was done to investigate the biological affinity. For this purpose, gene flow and population diversity present within the group, as well as between the study sample (N = 63) and other modern and historic population groups from southern, central and east Africa were explored. Available comparative groups included the historic Khoe-San from Riet River (N = 31), the Sotho-Tswana from southern Africa (N = 61), the Basuku from central Africa (N = 66) and the Bahutu (N = 53) and Teita (N = 24) from east Africa. Ten craniometric variables were selected and used to perform population structure analysis based on model bound quantitative genetics and multiple discriminant function analysis (MDA). Quantitative genetic distances revealed that the Khoe-San sample was closest to the Riet River group. Residual variance analysis performed on two-sample subsets of the Khoe-San group (Cape KS and Various KS) showed a higher level of heterogeneity in the Cape KS than seen in the Khoe-San from various other areas in southern Africa. MDA revealed that Khoe-San intra-sample variance is relatively high, with 44% of the sample (sexes pooled) classified into the Riet River group. The remaining individuals were classified (in decreasing order) into Bahutu (24%), Basuku (24%) and Sotho-Tswana (8%). Although the Khoe-San specimens are closest to the Riet River group, they are clearly not homogenous. Their high level of phenotypic diversity most likely originated from a complex population history involving many group interactions driven by social and political marginalization.