A rapid and sensitive analytical method based on high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) has been developed for the determination of felbinac in rat plasma, bile, urine, feces and tissue. Sample preparation involved liquid-liquid extraction with ethyl ether-dichloromethane (60:40, v/v). Chromatography of felbinac and the internal standard probenecid was performed within 2min on a Venusil MP C(18) column (100mmx4.6mm i.d., 5microm) with a mobile phase consisting of acetonitrile-5mM ammonium acetate containing 0.1% formic acid (pH 3.0) (80:20, v/v) at a flow rate of 1.2ml/min. Detection by electrospray negative ionization mass spectrometry and multiple-reaction monitoring of the transitions of felbinac at m/z 211.1-->167.0 and of probenecid at m/z 283.9-->239.9 was linear over the concentration range 5-5000ng/ml with a lower limit of quantitation of 5ng/ml using a sample volume of only 50microl. Intra- and inter-day precisions (as relative standard deviation, R.S.D.) were < or =7.3% and < or =6.4%, respectively, and accuracy (as relative error, R.E.) was in the range -2.1 to 7.4%. Recoveries and matrix effects were satisfactory in all the biological matrices examined. The method was applied to a preclinical pharmacokinetic study in rat involving a single intravenous injection of felbinac trometamol.