The interface of high-resolution capillary separation methods to time-of-flight mass spectrometry (TOF-MS) has generated considerable interest since TOF can provide the rapid and sensitive detection required by high resolution separations. In recent years, our laboratory has developed a variety of high-resolution capillary separation methods interfaced to TOF-MS via an ion trap storage/reflectron time-of-flight (IT/reTOF) instrument. Using this hybrid configuration, detection of fast separations at very low detection levels has been successfully performed for on-line separations of peptides and protein digests using electrospray ionization. In this report, we review the current status in our laboratory of interfacing high-performance liquid chromatography, capillary electrophoresis, and capillary electrochromatography to an IT/reTOF-MS instrument and various applications that have been developed involving this technology.