The permeation of ten model drugs through silicone membrane was analysed to investigate the effect of the presence of a biosurfactant obtained from corn steep liquor. The ten selected pharmaceutical compounds were chosen to include a diverse range of physicochemical properties, such as variable hydrophobicities, pKa’s, molecular masses and degrees of ionisation. When compared with compound permeation alone, the additional inclusion of biosurfactant in the donor phase altered the rate and extent of permeation. It significantly enhanced permeation for five of the compounds, whereas it decreased permeation for four of the compounds and remained approximately the same for the tenth compound. These effects were observed at both biosurfactant concentrations considered, namely 0.005 mg/mL, i.e. below the critical micellar concentration (CMC) and 0.500 mg/mL, i.e. above the CMC of the biosurfactant. Upon analysing permeation change with respect to physicochemical properties of the compounds, it was determined that compounds with a relative molecular mass below 200 resulted in an increase in permeation with biosurfactant present, and those above 200 resulted in a decrease in permeation with biosurfactant present. This effect was therefore attributed to the formation of a drug-biosurfactant interaction that enhanced permeation of smaller compounds, yet retarded permeation for those with a higher molecular mass. These in vitro findings can be considered an indication of potential novel formulation options that incorporate biosurfactant to create transdermal products that have bespoke permeation profiles.