DRUG FORMULATIONS Application of Design Space, Uncertainty, and Risk Profile Strategies to the Development and Validation of UPLC Method for the Characterization of Four Authorized Phosphodiesterase Type 5 Inhibitors to Combat Counterfeit Drugs Yassine Hameda Benchekroun, 1 Miloud El Karbane, 2 Bouchaib Ihssane, 1 Hasnaa Haidara, 1 Mohamed Azougagh, 3 and Taoufiq Saffaj 1 1 Universite ´ Sidi Mohamed Ben Abdellah, Faculte ´ des Sciences et Techniques, Laboratoire de Chimie organique Applique ´ e, Fe ` s, Maroc, 2 Universite ´ Mohamed V, Faculte ´ de Medicine et de Pharmacie, Laboratoire de Chimie Analytique et Bromatologie, Rabat, Maroc, 3 Universite ´ Mohamed V, Ecole Normale Supe ´ rieure de l’Enseignement Technique, Rabat, Maroc Corresponding author’s e-mail: yassine.benchekroun@usmba.ac.ma Abstract Background: Counterfeit medicines are an increasing scourge that are difficult to identify and they have become industrialized and widespread through highly organized illegal channels. Objective: This research aims to develop a robust method to determine four phosphodiesterase type-5 inhibitors in counterfeit drugs based on ultra-performance liquid chromatography. Method: Experimental design methodology (DOE) and design space (DS) recommended by ICH Q8 were used side-by-side in the development phase to define the optimal parameters as well as the robustness of the chromatographic method. Moreover, both the uncertainty and risk profile derived from the b-content and c-confidence tolerance interval were investigated during the validation phase to examine the performance of this method. Results: Successful chromatographic results, in a high resolution between the four active ingredients and an optimal analysis time of less than 1.6 min, were achieved at the end of the optimization phase. In addition, validation results show a low risk of future measurements outside acceptance limits set at 5%. Conclusions: Our procedure was successfully applied in the routine phase to identify 23 illicit formulations of an erectile dysfunction drug. Highlights: An efficient method for the characterization of 4 authorized phosphodiesterase in less than 1.6 min was established. A DS approach was applied to test the performance of this analytical method during analytical development. A risk profile was then carried out to approve the validity of the analytical method through the uncertainty profile approach. Received: 14 March 2019; Revised: 5 August 2019; Accepted: 24 September 2019 V C AOAC INTERNATIONAL 2020. All rights reserved. For permissions, please email: journals.permissions@oup.com 715 Journal of AOAC INTERNATIONAL, 103(3), 2020, 715–724 doi: 10.1093/jaocint/qsz006 Advance Access Publication Date: 19 April 2020 Article Downloaded from https://academic.oup.com/jaoac/article/103/3/715/5822176 by guest on 03 December 2022