The ex vivo permeation of a model peptide, tryptophan-leucine (Trp-Leu), was studied using two different regions of pig oral mucosa, the hard palate and the cheek. In order to increase the mucosal absorption of Trp-Leu, a lipophilic derivative was synthesized by acylation of the N-terminal amino group of Trp-Leu with myristic acid. The purified Trp-Leu derivative (Myr-Trp-Leu) was more lipophilic than the parent Trp-Leu as observed by HPTLC (Rf's values of 0.41 and 0.81, respectively). Measurement of partition coefficients in n-octanol/phosphate buffer pH 7.4, gave Kp values of −0.68 and 1.04 for Trp-Leu and Myr-Trp-Leu, respectively. The native Trp-Leu was unable to pass through the keratinized layer of palatal mucosa, and after 24 h only 12% had passed through the buccal mucosa to the receptor compartment. The higher lipophilicity of the acylated peptide, meant that it was not easily transported across the oral mucosal barrier but accumulated in the tissue, founding 25 and 70% of the original amount in the palatal and buccal mucosae, respectively. Both, Trp-Leu and Myr-Trp-Leu were found to be stable in palatal and buccal mucosae.