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A novel and reproducible isocratic normal phase liquid chromatographic method was developed for the quantitative determination of 10 stereoisomers of Nebivolol in pharmaceutical bulk drugs and dosage forms. The method was developed using an amylose-based chiral stationary phase, Chiralpak AD-3 (250 × 4.6 mm, 3 μm) column with mobile phase containing  n -hexane–ethanol–isopropanol–diethanolamine in the ratio 42:45:13:0.1 (v/v/v/v). The eluted compounds were monitored at 280 nm. Ten stereoisomers of Nebivolol were well separated with resolution &gt;2.0 for all pair of components. The developed method was validated as per International Conference on Harmonization (ICH) guidelines with respect to specificity, linearity ( R  2  value &gt;0.999), limit of detection, limit of quantification, accuracy (recovery range 95.8–103.2%), precision (relative standard deviation, RSD, &lt;2.5%) and robustness. Nebivolol sample solutions were found to be stable when characterized over a period of 48 h. Forced degradation studies were also performed to demonstrate the stability-indicating power of the developed HPLC method. The method was found to be rugged and robust.

A Validated Chiral LC Method for Enantiomeric Separation of Nebivolol Stereoisomers in Bulk Drugs and Dosage Forms on Amylose-Based Stationary Phase