¹Department of Pharmaceutical Chemistry, Bharati Vidyapeeth College of Pharmacy, Shivaji University, Kolhapur – 416013 DOI : 10.12991/mpj.2017.17 The estimation of individual plasma protein in free and bound form with analytes has importance in pharmacokinetics study. Albumins and globulins are most abundantly found in plasma and plays crucial role in plasma protein binding. The present communication deals with the development of gel permeation chromatographic method for the estimation of plasma protein binding studies for ketoprofen, tapentadol and furaltadone. The method was developed using WATER system with Ultrahydrogel column (7.8 x 300 mm i.d.), refractive index detector (1.00 to 1.75 RIU) and Rheodyne injection valve fitted with a 20μl sample loop using 0.1% sodium nitrate as a mobile phase. The method was developed by studying binding above drugs with most of plasma proteins. These complexes have shown linearity in range of 100 to 300μg/ml of these drugs. The developed method has been validated using USFDA guidelines. The developed bioanalytical method is accurate, precise, and selective and sensitive for quantification of plasma proteins bound drugs. The further optimization of method using other standard plasma proteins will explore its applicability in pharmacokinetic and biopharmaceutical studies of most of drugs. Keywords : Bovine serum albumin; gel permeation chromatography; plasma protein binding; human serum albumin