Chiral separation and quantitation of dorzolamide hydrochloride enantiomers by high-performance liquid chromatography

✍ Scribed by Mojtaba Shamsipur; Rouhollah Heydari; Elahe Sadat Jamasbi; Assem Abdollahpour

• Publisher: John Wiley and Sons
• Year: 2010
• Tongue: English
• Weight: 186 KB
• Volume: 33
• Category: Article
• ISSN: 1615-9306
• DOI: 10.1002/jssc.201000127

✦ Synopsis

Abstract

A direct HPLC method for chiral separation of dorzolamide hydrochloride (4__S__,6__S__) and its enantiomer (4__R__,6__R__) was developed. Dorzolamide (4__S__,6__S__) and its antipode were separated on a chiral‐α~1~‐acid glycoprotein column (150×4.0 mm, 5 μm). The influences of pH, temperature, flow rate, buffer concentration, and organic modifiers of the mobile phase on the retention and enantioselectivity were evaluated. The mobile phase consisted of an ammonium acetate buffer of pH 7.0. The method was validated for linearity, repeatability, accuracy, LOD, and LOQ. Calibration curves were constructed in the range of 0.5–10 μg/mL for dorzolamide (4__S__,6__S__) and 0.2–5 μg/mL for its enantiomer (4__R__,6__R__). Repeatability (n=6) showed less than 2% RSD. LOD and LOQ of the two enantiomers were found to be 0.2 and 0.5 for dorzolamide (4__S__,6__S__), 0.05 and 0.2 for its enantiomer (4__R__,6__R__), respectively. The proposed method was applied to the determination of dorzolamide enantiomer (4__R__,6__R__) in a raw material and two different eye drop samples.