Mycophenolic acid (MPA) is the active moiety of mycophenoate mofetil (MMF), an ester prodrug widely used as an immunosuppressant. Therapeutic drug monitoring (TDM) of MPA is becoming mandatory for transplant patients received MMF therapy in the routine clinical practice because of large individual variability, dose-related toxicity and the risk of acute rejection. In this study, a rapid, sensitive and selective LC-MS/MS method was developed and validated for the quantitative analysis of total and free MPA in plasma and in saliva that uses one identical liquid chromatographic and mass spectrometric condition. Following protein precipitation for total and saliva MPA, and ultrafiltration for free MPA, chromatographic separation was performed on an Allure PFP Propyl analytical column (100 x 2.1 mm, 5 microm, RESTEK Co., Bellefonte, PA, USA) with 0.1% formic acid in acetonitrile and 0.1% formic acid in water (45:55, v/v) as the mobile phases. The compounds were quantified by positive electrospray ionization tandem mass spectrometry. Selectivity, linearity, accuracy, precision, recovery, matrix effect, and saliva stability were evaluated during method validation. The validated method was applied to a pharmacokinetic study of MPA after an oral administration of a single 1000 mg of MMF to eight healthy male volunteers and 750 mg bid of MMF to nine renal transplant patients.