Development and Validation of a Robust LC-MS-MS Method for the Quantification of Amlodipine in Human Plasma
DOI:
https://doi.org/10.48047/HM.10.2.2024.1331-1341Keywords:
Amlodipine; LC-MS-MS, Method development, Validation, Human plasmaAbstract
The primary objective of this study was to develop a simple, precise, rapid, environmentally friendly, and highly accurate method for the simultaneous quantification of amlodipine in human plasma. Optimal peak resolution was achieved using an Intersil ODS gum C18 column (4.6 × 50 mm, 3.5 μm). Chromatographic separation was performed in gradient mode with a mobile phase comprising acetonitrile, water, and formic acid. The mobile phase was delivered at a flow rate of 0.500 mL/min into the ESI (Electrospray Ionization) chamber of the mass spectrometer. The total runtime for chromatographic analysis was 8.5 minutes, with amlodipine eluting at approximately 5.25 minutes. A linear calibration curve was established in the range of 1 to 130 ng/mL for amlodipine in human plasma. The method's efficiency was demonstrated by the mean percentage recoveries at various quality control levels: 104.1% at Low Quality Control (LQC), 105.0% at Medium Quality Control (MQC), and 108.7% at High Quality Control (HQC). The Lower Limit of Quantification (LLOQ) was determined to be 1.01 ng/mL, with a relative standard deviation (RSD) of less than 20%. The accuracy of the LLOQ was evaluated across different biological matrices, showing a value of 100.5% for amlodipine, which is well within the acceptable range of 80.00% to 120.00% across seven different plasma batches. The method was developed as a rapid, robust approach for the quantification of amlodipine in human plasma and was validated rigorously following the guidelines set forth by the International Council for Harmonization (ICH). The results confirm that this method is both reliable and efficient, making it suitable for routine analysis of amlodipine in human plasma samples.
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