Abstract: Objective To develop a specific and sensitive method for the determination of probucol in human plasma using HPLC-MS/MS technique. Methods Probucol were extracted from plasma by ethyl ether:dichloromethane (1:1, V/V), with physcion as an internal standard. The analytes went through the column of ultimate CN (50 mm×4.6 mm, 5 μm) with mobile phase acetonitrile:water:ammonia water=97:3:0.05 (adjusted pH=7.2 with formic acid). Probucol was analyzed with a negative mode. The ion pairs being detected were 515.5→236.1 (probucol) and 283.0→239.9 physcion, respectively. Results The established method was able to determine probucol in human plasma over the range of 2.5-6000 ng/mL, with a method recovery ranging from 93.02% to 104.12%. The intra and inter day variances were below 4.67% and 5.72%. Conclusion The HPLC-MS/MS method for analyzing probucol was validated. It is sensitive and suitable for pharmacokinetic studies of probucol.