RP-HPLC method development and validation for novel synthesized polyethylene glycol conjugated isotretinoin prodrug

Orally administered Isotretinoin is a highly effective medication for treating cystic acne. The current study is to synthesize Isotretinoin prodrug via an ester linkage to polyethylene glycol (PEG) 4000, with DCC (dicyclohexyl carbodimide) as a coupling agent and DMAP (dimethylamino pyridine) as a catalyst. FT-IR, DSC and 1H NMR were used to characterize the synthesied prodrug. RP-HPLC method was developed and validated for prodrug. Thermo C18 (Hypersil BDS) column (250 mm x 4.6 mm, 5 mu m) was used for chromatographic separation with acetonitrile: water (95:05% v/v) as a mobile phase. UV detection was done at 344 nm; flow rate was 1mLmin(-1) and an column temperature of 35degree celsius. Total runtime of the analytical run was 15 min. High selectivity and good precision features were found in the method validation results, which were examined at six concentration levels. In intra- and inter-assay precision experiment, the RSD was less than 2.0%. The recovery of isotretinoin, which was examined at three different degrees of fortification, ranged from 99.06% to 100.42% and linearity was found for a range of 0. 2 to 100 mu g mL(-1). The calibration curve was linear over the 0.5-100 mu g mL(-1) concentration range. Photolytic stability of prodrug was performed under UV light. There was successfully improves the photolytic stability of Isotretinoin prodrug as compare to Isotretinoin.