Abstract: In alkaline solution, vanillin shows strong sensitivity enhancement effect to chemiluiminesecnce product by luminol- K3Fe ( CN) 6system. According to this phenomenon, a kind of by high performance liquid chromatography ( HPLC) coupled separation column after the determination of vanillin chemiluminescence method. This method was based on the C18 column, acetonitrile and water (50∶50, V/V) as mobile phase, under the optimal conditions, the chemiluminescence intensity was correlated linearly with concentration of vanillin in the rang of 7. 0 × 10-7~ 4. 0 × 10-5mol·L-1, and the determination limit was 2. 0 × 10-7mol·L-1 ( 3 σ) , its relative standard deviation was 3.9% (c =4.0 ×10-6mol/L, n =9) . This method had been applied to the determination of vanillin in food samples, the analysis results agreed well those from HPLC-UV method.