The online hyphenation of chip-based high-performance liquid chromatography (chip-HPLC) with ion mobility spectrometry (IMS) via fully integrated electrospray emitters is introduced. A custom-built drift tube IMS with shifted potentials was developed in order to keep the IMS orifice electrically grounded, allowing for a robust coupling with chip-HPLC. Proof-of-concept studies with the newly developed analytical setup revealed the suitability of IMS as a promising and powerful detection concept for chip-based separation techniques. Comparison of IMS with fluorescence detection and electrospray ionization-mass spectrometry (ESI-MS) allowed a more detailed characterization of the IMS as a new detection method for chip-HPLC. Moreover, the analysis of a mixture consisting of three isobaric antidepressants demonstrated the performance of chip-HPLC/IMS as a miniaturized two-dimensional separation technique.