Aim: The present investigation shows confirmatory evidence for the complex formation of Rosuvastatin with a fraction of Clarified Butter. Materials and Methods: Fractions of Clarified Butter prepared according to thermal behavior at different temperatures (30°C, 40°C, 50°C). Rosuvastatin biform complex with Clarified Butter in the ratio (1:1) and (1:2) w/w were prepared. Physicochemical properties of all fractions of Clarified Butter was determined as per Indian Pharmacopoeia. The fractions were subject to Fourier-transform Infraredspectroscopy (FTIR), Differential Scanning Calorimeters (DSC) and in-vitro release studies (Simulated gastric fluid, pH 1.2 for 2 h and simulated intestinal fluid, pH 6.8 for 6 h). Results: Based on these investigations; the optimized ratio (1:1) w/w were further characterized for DSC and FTIR studies. Where Rosuvastatin exhibited a strong carbonyl band at 1543.05 cm−1, all fractions showed the disappearance of strong carbonyl band at 1543.05 cm−1, spectra of biform complex shifted to higher frequency 1747.51 cm−1 (13.29 % increase), that confirmed carboxylic and hydroxyl group of both saturated and unsaturated fatty acid present in Clarified Butter. From in vitro release study the highest percent release i.e. 99.23±0.08% of Rosuvastatin was obtained from 1:1 biform complex. DSC thermogram revealed the formation of eutectic mixture by lowering the peak of pure Clarified Butter (67.4°C) to 58.10°C due to the formation of biform complex. Conclusion: Thus, Clarified Butter and its fraction illustrated enhanced bioavailability of Rosuvastatin.