Objective: The main objective of this study was to assess the impact of inter-occasion variability (IOV) on Trospium plasma concentration level from typical crossover pharmacokinetic study using non-compartment model analysis. Methods: An open, randomized, fasting, single-dose, two way crossover reference replicate study was performed with 36 healthy, non-smoking, male subjects. Plasma concentration of Trospium was estimated. The influence of inter-occasion on Trospium pharmacokinetics was evaluated using non-compartment model analysis. Results: Results from the non-compartment analyses showed that inter-occasion variability as measured by coefficient of variation was found more than 30% for pharmacokinetic parameters Cmax, AUClast and Vd/F. In addition, IOV for Cmax was higher when compared with any other pharmacokinetic parameters. Conclusion: Overall, considerable variability associated with Trospium chloride pharmacokinetics between occasions was established. Trospium was well tolerated in the treated subjects and there was no serious adverse event noted in the entire study. Thus, major differences in obtained variability seen in this study are likely to be of only limited clinical significance.