In the present investigation, we have designed a gastroretentive drug delivery system of moxifloxacin for the pharmacokinetic profiling of this system by relating it with moxifloxacin controlled release drug delivery system. A fast, simple, precise, specific and sensitive high performance liquid chromatography (HPLC)-ultraviolet-visible (UV) method was developed and validated for the plasma sample analysis. The C18 column was deployed for moxifloxacin separation using the mixture of acetonitrile and 0.5% triethylamine solution. The calibration curve was linear for concentration range of 10–1000 µg/L. A pharmacokinetic study was performed using oral single dose of 5 mg/kg moxifloxacin in healthy white New Zealand rabbits (n = 6) by administrating gastroretentive and controlled release systems respectively. One compartment open model was applied for the estimation of pharmacokinetic parameters after oral administration. The controlled release system of moxifloxacin shows significantly lower value average steady state plasma concentration (884.33±5.27 µg/L) than gastroretentive system (1389.67±11.31 µg/L). The quick attainment of steady state level of drug in plasma indicates higher value absorption rate constant and lower absorption half-life for gastroretentive system. The area under curve for controlled and gastroretentive system were found to be 9.57±0.53 and 27.34±1.32 mg.h/L respectively. The results concluded that gastroretentive system have 2.86 folds greater relative bioavailability and double value of mean residence time in comparison to controlled release system.