Characterızatıon of vasoactıve ıntestınal polypeptıde (VIP) and pıtuıtary adenylate cyclase-actıvatıng polypeptıde (pacap) wıth Lc/MS-Ms method

Neuropeptides are peptides used for communication by neurons. Vasoactive intestinal polypeptide (VIP) and pituitary adenylate cyclase-activating polypeptide (PACAP) are obtained from the hypothalamus region of the brain, glucagon/secretin family of polypeptides. VIP and PACAP are quite similar to each other in terms of amino acid sequences. In this thesis, the analytical characterization of VIP and PACAP-38 by sensitive Liquid Chromatography-Mass Spectrometry (LC-MS / MS) method have been carried out. For the analysis of both peptides, the parameters for chromatography (C8 and C18 separation columns, organic solvent and acid content of the mobile phase, flow rate etc.) and mass spectrometry (observation of basic ions and fragments of peptides, fragmentor voltage, fragmentation energy, etc.) were examined. The sources of the adsorption problem of the peptides were investigated and the factors inhibiting adsorption were tested in LC-MS / MS analysis. In the analysis, 665,9 (M+5) m/z base ion and 799,5 and 770,7 m/z fragment ions were observed for VIP, while 504,3 (M+9), 567,3 (M+8), 648,3 (M+7) and 755,9 (M+6) m/z basic ions for PACAP-38 were investigated. In addition, the effects of albumin solution and surfactants on the analysis were investigated as inhibitor of adsorption of peptides to the surface.