VALIDATED RP-HPLC METHOD DEVELOPMENT FOR THE SIMULTANEOUS ESTIMATION OF ACETYLCYSTEINE AND ACEBROFYLLINE IN CAPSULE FORMULATION

Abstract

A new simple, precise, rapid and accurate reverse phase high performance liquid chromatographic method had been developed for the simultaneous estimation of Acetylcysteine (ACST) and Acebrofylline (ACBF) in capsule dosage form. The chromatographic separation was achieved on a Hypersil BDS, C18, 100 x 4.6 mm, 5mm particle size column was used with PDA detector by using mobile phase containing mixture of 0.02M Potassium dihydrogen orthophosphate (KH₂PO₄) buffer : acetonitrile (90:10 % v/v pH 3.2) was used. . The flow rate was 0.9 ml / min and effluents were monitored at 260 nm. Chromatogram showed two main peaks corresponding to Acetylcysteine and Acebrofylline at retention times 2.365 and 5.505 min respectively. The method was liniear over the concentration range of 150-900µg/ml for Acetylcysteine and 25-150 µg/ml for Acebrofylline respectively. The developed method was validated in accordance to ICH guidelines.

Key words: Acetylcysteine, Acebrofylline, RP-HPLC, Validation, ICH, Acetonitrile