Toads skins secretions are sources of complex mixtures of bioactive compounds such as proteins and peptides. Rhinella jimi specie is a common toad in Brazilian northeast with little known studies. The fractional factorial design was used to study the best method for extraction of the proteins from R. jimi paratoid gland secretions founding Tris/HCl pH 7.2 with vortex stirring for 5 min as the optimal parameters. A characterization of extracts and brute venom by FTIR spectroscopy showed that the extraction successfully separate the proteins from the rest of the secretion. A SEC-HPLC analysis compared the proteome between two cities, showing 464 and 171 kDa for Picos city and 131 and 11 kDa for Teresina city, and also outside of the size exclusion range a peptide like mass composition was observed. With all these preliminary data is expected the future studies to identify the proteins and peptides as also evaluate their biological activities.