Abstract Aim: The aim of the present investigation is to study RP-HPLC Method Development for Estimation of Various Pharmacokinetic Parameters of Optimized Formulation of Ganciclovir. Material & Methods: In the preformulation study & on the basis of previous formulation, solubility profile and release rate characteristics were found to be good in the formulations with PEG-8000, PVP-17, Poloxamer-188and Urea. So, solid dispersions were tried in these combinations. Healthy rabbits of either sex (weighing 1.5 – 2.5 kg) were fasted overnight. GCR and its solid dispersions were administered at dose equivalent to 2.6mg/kg of GCR. Each product was repeated 4 times (n = 4). The in vivo experiments were conducted as a crossover study. GCR in serum samples were estimated according to High Performance Liquid Chromatographic (HPLC) method. Results: The best formulation (among F-1 to F-6) i.e. F-5 was exposed to in vivo examinations in rabbits. An overlay spectrum of GCR indicated λ max at 250.2 nm. The simultaneous estimation of GCR was done by RP-HPLC. The retention time of GCR was found to be 2.578 min the asymmetric factor was within limits. After a single dose of formulation F-5 (GCR - 2.6mg/kg), the symmetrical mean C max values of formulation F-5 (47.37±1.52 µg/mL for GCR), was obviously higher than that of pure drugs {GCR (P<0.05), which was 21.26±0.03 µg/mL. Cmax of SDs (F-5) was 2.22 times more than GCR. The T max values of the formulations F-5 was equivalent to the pure drug. The AUC (0-8h) values of the formulation (GCR 266.6±5.65 µg.h/mL) was obviously higher than those of the pure drugs (GCR 132.65±1.48 µg.h/mL). Conclusion: The in vitro drug release from the best SD combination (F-5) was compared with marketed sample was revealed that the drug release from the SDs was on par equivalent with the marketed dosage form.