Box-Behnken response surface modeling assisted enantiomeric resolution of some racemic β-blockers using HPTLC and β-cyclodextrin as chiral mobile phase additive: Application to check the enantiomeric purity of betaxolol.

Abstract

Stereospecific separation method of (±) betaxolol, (±) carvedilol, and (±) sotalol using High Performance Thin Layer Chromatography (HPTLC) and β-cyclodextrin as chiral selector has been developed and validated. The Box-Behnken surface response design was selected for optimizing the operating variables based on 15 trials design. The optimized method involves separation on Fluka HPTLC silica gel plates 60F254 (20×10cm) using acetonitrile-methanol-acetic acid-water (3.4:3.6:0.18:1v/v) as a mobile phase containing 0.57mM β-cyclodextrin. Densitometric measurements were made at 220nm for betaxolol and sotalol or at 245nm for carvedilol. Maximum separation of the enantiomers of the three drugs was obtained by optimizing concentration of chiral selector, the mobile phase composition including acetonitrile amount in the organic part of the mobile phase and the volume of acetic acid added. The proposed method enables estimation of (-) and (+) enantiomers of betaxolol in drug substance and in various pharmaceuticals. The detection limit of betaxolol was 0.15 and 0.13μg band-1 for (-) and (+) enantiomers, respectively. The detection limits were found to be 0.2 and 0.3μg band-1 for carvedilol and sotalol, respectively, as racemate. In addition, the proposed method was applied in checking the enantiomeric purity of (-) BET in the presence of (+) BET at 1% level where the inactive (+) enantiomer was quantified with good accuracy and precision at 1% level in the active (-) enantiomer.