Optimization and modeling of a green dual detected RP-HPLC method by UV and fluorescence detectors using two level full factorial design for simultaneous determination of sofosbuvir and ledipasvir: Application to average content and uniformity of dosage unit testing

Publication date: June 2019Source: Microchemical Journal, Volume 147Author(s): Hanan I. EL-Shorbagy, Fawzi Elsebaei, Sherin F. Hammad, Amina M. El-BrashyAbstractA simple, rapid, selective, linear, precise, accurate and eco-friendly RP-HPLC-UV-Fluorescence method was developed by 23 full factorial design and validated for simultaneous assay of sofosbuvir and ledipasvir mixture in tablet dosage form within 7 min without interference of tablet excipients. Isocratic elution at a flow rate of 1.2 mL min−1 was employed on C18 (250 mm × 4.6 mm, 5  μm in particle size) at ambient temperature. The column equipped with UV detector then fluorescence detector. The fluorescence detector is an optional extra tool for a more sensitive analysis of LED. UV detector can be used for both drugs. UV detector was set at 261 nm and switched online at 5 min to 333 nm. On the other hand, the fluorescence detector was set at an emission wavelength of 404 nm with excitation at 333 nm. The mobile phase consisted of acetonitrile:methanol:0.01% triethylamine adjusted to pH 3 by glacial acetic acid [35: 35:30] (v/v/v). After injection of 30 μL sample, the retention time for sofosbuvir by UV detector was 3.353 ± 0.017 min. While for ledipasvir, it was 6.102 ± 0.006 min by UV detector and 6.255 ± 0.004 min by florescence detector. Using the proposed method, calibration graphs were found linear for sofosbuvir over the concentration range of 1–4...
Source: Microchemical Journal - Category: Chemistry Source Type: research