Simultaneous estimation of nebivolol hydrochloride and amlodipine besylate in human plasma employing an innovative HPLC chromatographic method

Background

This study developed and validated a simple, robust, and cost-effective RP-HPLC bioanalytical method for the determination of nebivolol hydrochloride (NBH) and amlodipine besylate (AMB) in human plasma. Briefly, NBH and AMB were extracted from plasma through protein precipitation using 5% formic acid and acetonitrile. Chromatographic separation was achieved using an Inertsil ODS-3 V column (150 mm × 4.6 mm, 5 μm) with a mobile phase composed of acetonitrile and buffer (40:60, v/v). The analysis was conducted using UV detection at 215 nm.

Results

The bioanalytical method demonstrated linearity for NBH (4.50–180.12 μg/mL) and AMB (3.50–140.06 μg/mL). It exhibited good selectivity and sensitivity, with LLOQ responses within ≤ 20% of the analyte signal. Accuracy and precision were within acceptable limits. The extraction recovery from human plasma showed a CV (%) of 1.15% for NBH and 1.35% for AMB, indicating consistent recovery rates. Stability studies on drug-spiked human plasma at LQC and HQC levels confirmed the stability of the drugs under various conditions.

Conclusion

The present bioanalytical method successfully quantified NBH and AMB simultaneously in plasma samples. It demonstrated suitability, supported by high recovery rates and low relative standard deviations. With its proven linearity, accuracy, and precision, this technique is well suited for drug identification in plasma samples.