Calixarenes, following cyclodextrines and crown ethers, are the third generation of supramolecules used in HPLC as stationary phases. They consist of phenol units linked via methylene bridges and can also form inclusion complexes like the other host supramolecules. The resulting interactions influence the retention factors and improve the selectivity of the solutes. Additionally, modification of the calixarenes, for instance by varying the ring size, substitutents, conformations and pKa values, enable a more enhanced interaction spectrum and can improve the specificity for guest molecules. The application of calixarenes in chromatography also includes medical and environmental applications, preparative chemistry as well as the rapidly developing area of supramolecular chemistry. Taking the possibilities and the growing interests of calixarenes into account, the aim of the review is to summarise the application possibilities and interactions of calixarenes as stationary phase in HPLC.