Novel Stability-Indicating High-Performance Thin Layer Chromatography (HPTLC) Method Development and Validation for Estimation of Daclatasvir Dihydrochloride in Pharmaceutical Dosage Form

Abstract

Daclatasvir dihydrochloride (DTDH) is a direct-acting antiviral agent used against the hepatitis C Virus. Different analytical methods were reported for the determination of DTDH in pharmaceutical dosage form, including spectrophotometry, high-performance liquid chromatography (HPLC), and stability- indicating HPLC method but to date, no HPTLC method has been reported. A novel, simple, sensitive, selective, precise, and accurate stability-indicating high-performance thin-layer chromatographic (HPTLC) method has been developed for the quantification of Daclatasvir dihydrochloride (DTDH) in pharmaceutical formulation. Chromatography separation was carried out on pre-coated silica gel G60 F254 HPTLC plates using chloroform: methanol (9:1, v/v) as the mobile phase. Scanning and densitometric analysis were done at 316 nm. The band of DTDH were obtain at RF= 0.48±0.02. The calibration curve was plotted in the concentration range of 400– 900 ng/band and the method was found linear with r = 0.995. The LOD and LOQ were found to be 95 ng/band and 313.5 ng/band respectively. The percentage recovery was found to be 97.31-101.81 %. The developed was able to separate peaks of all the degradation products formed in ICH-prescribed stress conditions with a significant difference in their RF values. The developed method was validated according to ICH guidelines and results were found to be within acceptance criteria. The developed stability-indicating method successfully evaluated the DTDH in pharmaceutical tablet dosage form and assay results showed good recovery when statistically compared with the reversed-phase high-performance liquid chromatography (RP-HPLC) method.