In current scenario, treatment of any disease depends upon two major factors i.e. patient compliance and effective dosage regimen. The effective dose delivered by a dosage form to a patient depends on various parameters, which can be assessed by an effective and economic analytical method. In the present study a precise analytical method for estimating the combination of immunosuppressant drugs mycophenolate mofetil (MMF), tacrolimus (TAC) and prednisolone through RP-HPLC was developed. The mobile phase contained a mixture of acetonitrile and 0.35% triethylamine (pH 4.2) with orthophosphoric acid (70:30). As per ICH guidelines the optimized RP-HPLC method was validated with respect to linearity, limit of detection (LOD), limit of quantitation (LOQ), accuracy, precision, repeatability, robustness, ruggedness. The accuracy of the method was determined in terms of % recovery of the standard. The obtained test results were compared with that of the standard drug. The results of the recovery study were found to be within the acceptance criteria (96.93- 103.99%), which indicated a good degree of sensitivity of the developed method in detection of analytes in a sample.