A self-assembled bilayer lipid-like membrane (BLM) supported on glassy carbon electrode (GCE) was fabricated using 5,5-ditetradecyl-2-(2-trimethyl-ammonioethyl)-1,3-dioxane bromide (DTDB) for epinephrine (EP) determination in the presence of ascorbic acid (AA). This modified electrode (DTDB/GCE) has strong membrane adsorption accumulation and electrocatalytic ability toward EP and AA. The oxidation of EP was controlled by double step adsorption accumulation process of the DTDB-BLM. The parameters of fitted Langmuir isotherm Gamma(max), B-ADS, and DeltaG(ADS) values were determined as 1.0 x 10(-11) mol cm(-2), 2.04 x 10(6) dm(3) mol(-1), and -45.17 kJ mol(-1) for the fist step for EP concentration less than 1 mM, and 4.92 x 10(-11) mol cm(-2), 7.35 x 10(4) dm(3) mol(-1), and -37.1 kJ mol(-1) for the second step for EP concentration higher than 1 muM. The DPV peaks for EP and AA oxidations were appeared at 0.220 and 0.085 V versus SCE, respectively, allowing the determination of EP in the presence of high concentration of AA. The advantage of DTDB-BLM was demonstrated experimentally in comparison with other three BLMs, and attributed to the dioxane group as well as the suitable length of the carbon chain of DTDB molecule. The current response of the DTDB/GCE was fast and reproducible, suitable for the electrochemical sensing in flow-injection systems. A linear range of 1 x 10(-8) to 1 x 10(-4) M EP was preliminary obtained using a simple setup. (C) 2004 Elsevier Ltd. All rights reserved.