Mesoporous carbon containing polyurethane/clay membrane based sensors were produced for the rapid and sensitive electroanalytical determination of fat-soluble vitamin D2. Mesoporous carbon structures were prepared by the carbonization of bee pollens which were easily available in nature. These structures were combined with clay and polyurethane structures. The determination of vitamin D2 with modified electrodes was carried out by using square wave voltammetry in acetonitrile containing 0.1 M LiClO4. According to the response of modified electrode, a linear relationship was obtained between the anodic oxidation peak currents and concentrations of vitamin D2 in the linear concentration range (23 mu M - 844 mu M) with a low detection limit (0.73 mu M) was obtained. The sensitivity (4.65 mu A cm- 2 mu M- 1) and reproducibility (RSD%, 1.87 %) of the prepared novel sensor were found to be quite satisfactory. The proposed method and applicability of the electrode were tested in urine samples, achieving dramatic recovery values (99.99 %).