A new approach was applied for the development of a precise, simple, and economic analytical process for the accurate analysis of tolterodine tartrate (TOT) in its bulk and tablet using HSPiP- and quality by design (QbD)-assisted methods. The HSPiP program predicted several solvents and their right ratios for the mobile phase, followed by simulating the experimental solubility data in various predicted solvents. QbD was used to identify the impact of the composition and the mobile phase flow rate on the peak area and retention time. TOT was estimated using an Agilent TC C18 column employing an optimized mobile phase. The HSPiP shortened the solvent selection time with high reliability, whereas QbD identified critical factors. The optimized composition and process variables were used to develop an analytical method for TOT estimation. Various analytical validation parameters were estimated with constructed linearity of 5–30 μg/mL and a percent recovery yield value of 100.36%. To ensure the reliability of the optimized method, we estimated validation parameters (linearity, specificity, precision, accuracy, robustness, and ruggedness) to comply with the ICH guidelines. Considering the high recovery yield, good regression coefficient, low detection limit, and low noise ratio, the optimized method was accurate and precise with a high degree of specificity, rapid process, and reproducibility for the quantitative estimation of tolterodine from both oral analytes (I and II). The validated method was implemented for pharmacokinetic study in rats for quantitative estimation of the analytes with high accuracy, sensitivity, and reproducibility.