Experimental Design-Assisted HPTLC Method for Nirmatrelvir and Ritonavir Analysis in Pharmaceutical Dosage Forms

Ratna Musale1, Hiralben Mehta1*, Anas Jamsa2, Dilsar Gohil3 and Pooja Uttekar4

1*Assistant Professor, Department of Quality Assurance, Faculty of Pharmacy, Parul Institute of Pharmacy and Research, Parul University, Post: Limda, Vadodara, Gujrat-391760, India.
2Assistant Professor, Department of Pharmacology, Faculty of Pharmacy, Parul Institute of Pharmacy and Research, Parul University, Post: Limda, Vadodara, Gujrat-391760, India.
3Assistant Professor Sumandeep Vidyapeeth deemed to be University, Department of Pharmacy, Vadodara, Gujarat-391760, India.
4Research scholar, faculty of pharmacy, Parul Institute of Pharmacy and Research, Parul University, Post: Limda, Vadodara, Gujrat-391760, India.
1*heerm1990@gmail.com.

Received: 14th Sep, 2025; Revised: 20th Oct 2025; Accepted: 6th Nov, 2025; Available Online: 1st December, 2025

ABSTRACT

Background: This study aimed to develop and validate a High-Performance Thin Layer Chromatography (HPTLC) method utilizing a Design of Experiments (DOE) approach for simultaneous quantification of Nirmatrelvir (NIR) and Ritonavir (RITO) in pharmaceutical dosage forms.

Method: Pre-coated HPTLC aluminium plates with silica gel 60 F254 were employed as the stationary phase to optimize the mobile phase composition. Fractional factorial design (FFD) was applied to assess method robustness. The analytical procedure was validated following ICH Q2 (R2) guidelines.

Results: Detection was carried out at 260 nm for both NIR and RITO. Optimal separation was achieved using a mobile phase of ethyl acetate, n-hexane, and methanol in a ratio of 8:1.2:0.8 v/v/v. The Rf values observed were 0.46 for NIR and 0.78 for RITO. The method was validated for multiple parameters, with %RSD values falling within acceptable limits. FFD results indicated that all four factors significantly influenced the retention of NIR, whereas RITO was not significantly affected.

Conclusion: The developed HPTLC method is simple, rapid, selective, accurate, and precise. Statistical evaluation demonstrates reproducibility with no significant differences, indicating suitability for routine analysis of marketed formulations without prior separation.

Keywords: Nirmatrelvir, Ritonavir, HPTLC, DoE approach, Validation

How to cite this article: Musale R, Mehta H, Jamsa A, Gohil D, Uttekar P; Experimental Design-Assisted HPTLC Method for Nirmatrelvir and Ritonavir Analysis in Pharmaceutical Dosage Forms. Int J Drug Deliv Technol. 2026;16(1): 459-465. DOI: 10.25258/ijddt.16.1.48