Use of capillary electrophoresis-electrospray ionization mass spectrometry in the analysis of synthetic peptides

We have constructed a capillary electrophoresis (CE) system with UV detection and have successfully interfaced it to an electrospray ionization mass spectrometry (ES-MS) system. A synthesized fragment of heregulin-β (212–226) was thought to be a single component by re-injection into an HPLC system, but results from CE-UV-ES-MS indicated that a dehydration product was present in the desired peptide sample. A synthetic heregulin-α (177–241) was isolated by preparative HPLC, but re-injection on an analytical system indicated a tailing peak. CE-UV-ES-MS indicated a mixture whose two major components were of the same nominal molecular mass (within experimental error), suggesting the presence of an isomer or a deamidation product. The results show that CE-UV-ES-MS can be used as an orthogonal analytical technique to solve practical problems encountered in peptide synthesis laboratories.