In this paper, CE coupled with electrochemiluminesence (ECL) detection using a 76-mu m Pt disk as working electrode was developed for nicotine (NIC) determination. The major metabolite of NIC is cotinine (COT), which has a similar tertiary amine structure to NIC. However, there is a carbonyl group attached in the structure of COT, which leads to the great decrease in ECL response. In order to improve the ECL response of COT, NaBH4 was used for carbonyl reduction. After reduction, NIC and COT were separated and detected by CE-ECL. ECL response plotted with NIC concentration was linear between 5.0 x 10(-7) and 5.0 x 10(-5) mol/L (81-8100 mu g/L), with LOD of 5.0 x 10(-8) mol/L (8.1 mu g/L). The developed CE-ECL method was applied for NIC determination in urine and cigarette samples.