Development and Validation of a New Stability Indicating RP-HPLC Method for the Determination of Trifluridine
Sai Gnaneswari Aluri and Mukthinuthalapati Mathrusri Annapurna*
Department of Pharmaceutical Analysis, GITAM School of Pharmacy, Gandhi Institute of Technology and Management (Deemed to be University), Visakhapatnam, India
*Corresponding Author: Mukthinuthalapati Mathrusri Annapurna, Department of Pharmaceutical Analysis, GITAM School of Pharmacy, Gandhi Institute of Technology and Management (Deemed to be University), Visakhapatnam, India.
October 21, 2022; Published: November 30, 2022
Trifluridine is an anti-viral drug used for treatment and prevention of vaccinia viral infections on eye. Trifluridine is used to treat herpes viral infections as well as the inflammation of eye. The present study describes the development and validation of RP-HPLC method for the estimation of Trifluridine in ophthalmic preparations using Shimadzu Model CBM-20A/20 Alite HPLC system consisting of PDA (Photo diode array) detector. A mixture of Formic acid: Methanol (45:55, v/v) was used as mobile phase with flow rate 1.0 mL/min. The proposed method was developed using C18 Agilent column (250 mm × 4.60 mm, 5 μm) on an isocratic mode at room temperature with UV detection at 259 nm. Trifluridine obeys Beer-Lambert’s law over the concentration range 0.5-120 µg/mL with linear regression equation y = 40447x - 793.8 (R² = 0.9997). The method was validated as per ICH guidelines and showed excellent precision, robustness, accuracy, linearity, specificity and the system suitability parameters were within the acceptable criteria. The limit of detection (LOD) and limit of quantification (LOQ) values were found to be 0.1541 µg/mL and 0.4691 µg/mL respectively. Forced degradation studies were performed by exposing Trifluridine to stress conditions such as acidic hydrolysis, alkaline hydrolysis, thermal degradation and oxidation and the degradant products were separated successfully from the API.
Keywords: Trifluridine; Limit of Quantification (LOQ); Limit of Detection (LOD)
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