Pig ear skin membrane-covered glucose biosensor based on oxygen electrode has been assessed as a tool to evaluate glucose penetration through skin in-vitro. For this, glucose oxidase (GOx) was immobilised on oxygen electrode and covered with the skin membrane. Exposing this electrode to the solution of glucose resulted in glucose penetration though skin membrane, its oxidation catalysed by GOx, consumption of O-2 and decrease of the current of the oxygen electrode. By processing the biosensor responses to glucose, we found that glucose penetration through 250 pm thick skin membrane is slow; 90% of steady-state current response was reached in 32(+/- 22) min. Apparent diffusion coefficient for glucose in skin was found to be equal to 0.15( +/- 07)* 10(-6) cm(2) s(-1). This value is 45 times lower than glucose diffusion coefficient in water. Tape-stripping of stratum corneum (SC) allows considerably faster glucose penetration. The electrodes covered with tape-stripped skin reached 90% of steady-state current response in 5.0(+/- 2.7) min. The theoretical estimate of glucose flux through SC was considered exploiting four-pathway theory of transdermal penetration. Theoretical flux values were more that three orders lower than measured experimentally. This high discrepancy might indicate that glucose penetration through healthy human skin could be even slower, allowing much lower flux, than it was found in our study for skin membranes from pig ears.