The influence of the nonionic surfactant lauryl alcohol ethoxylate with 12 moles ethylene oxide (LAE-12OE) was evaluated on the Stratum corneum model biomembrane (SCMM) of shed snake skin (Bothrops jararaca and Spilotes pullatus) through the biophysical techniques Fourier transform Raman spectroscopy (FT-Raman) and Fourier transform infrared photoacoustic spectroscopy (PAS-FTIR). The surfactant was used in aqueous solutions above and below the critical micelle concentration (cmc), 50.0 and 0.21 g/L, respectively. The SCMM samples were pre-treated for periods of 8 h (whole SCMM) and for 12 h (SCMM after tape stripping procedure). The LAE-12OE did not promote increase in the hydration of the B. jararaca and S. pullatus SCMM but exhibited some action as far as lipid extraction.