In recent years, sequence-defined oligomers (SDOs) gained increasing interest due to their perfectly controlled molecular structure, thus providing defined properties. In order to tune the properties, different functionalities need to be incorporated into the oligomers and the chain tacticity needs to be controlled. Beside the synthesis of SDOs, suitable methods need to be found to analyze the molecular structure. In this work, oligomers exhibiting an alternating or block-wise sequence of side chain functionalities were analyzed using a hyphenation of ultra-high-performance liquid chromatography and electrospray ionization mass spectrometry enhanced by ion mobility separation (IMS). Moieties in the side chains were varied according to polarity and bulkiness. Moreover, chain tacticity was varied. Drift times in the IMS cell and the corresponding collision cross section (CCS) values were shown to be individual parameters allowing the identification of SDOs, even in the case that SDO structures only differ in sequence or tacticity of side chain functionalities. Thus, a library of CCS values was obtained as reference used for the analysis of complex mixtures of SDOs.