Metrological Evaluation of Metopimazine HPLC Assay: ISO-GUM and Monte Carlo Simulation Approaches

Background: Measurement uncertainty (MU) is a crucial parameter for ensuring the reliability of analytical methods and the validity of results, as required by ISO 17025:2017. Its estimation is particularly critical for quality control laboratories, where compliance decisions are based on a rigorous interpretation of uncertainties. Methods: In this study, we evaluated the uncertainty associated with an HPLC-UV method for the determination of Metopimazine (MPZ) in a pharmaceutical form, applying two complementary approaches: The ISO-GUM (Guide to the Expression of Uncertainty in Measurement) top-down approach and the Monte Carlo Simulation (MCS). Results: The results of both approaches showed excellent agreement, thus validating the robustness of the evaluation. The analysis of uncertainty sources revealed that the accuracy of the sample volume (V_Sample) and the calibration standard (C_x) were the dominant contributors, representing 39.9% and 36.2% of the total uncertainty, respectively. Combined, these two factors accounted for 76.1% of the variability, underscoring their critical impact on the assay's precision. The expanded uncertainty (k = 2, 95% confidence level) was determined to be (99.41 ± 0.69)%, reflecting the method's reproducibility. Conclusions: These results highlight the importance of rigorously controlling calibration standard preparation, sample volume, and repeatability conditions to optimize the reliability of the assay.