This review describes new analytical methodologies applicable to peptide purity validation and process control, The HPLC section is centered on the description of new stationary phases, which could offer better performance with respect to the well consolidated RP-HPLC techniques. In the chapter devoted to capillary electrophoresis the different options offered by zonal procedures are particularly stressed. The applications of either capillary electrophoresis in polymeric solutions, either isoelectric focusing as well as of isotacophoresis are subsequently discussed. The section devoted to the description of electrochromatography emphasizes, in particular, micellar electrokinetic capillary chromatography. Furthermore, future applications of packed electrochromatography are illustrated. The conclusion is dedicated to several hyphenated opportunities and future perspectives of peptide analysis, as micro total analytical systems.