Analytical Method Validation for the Quantification of Esomeprazole and Pantoprazole in Human Plasma Using RP-HPLC–MS/MS: Application to Pharmacokinetic Studies
Keywords:
Esomeprazole, Pantoprazole, RP-HPLC–MS/MS, PharmacokineticsAbstract
The accurate quantification of pharmaceuticals in human plasma is critical for pharmacokinetic and bioequivalence studies. This study presents the development and validation of an analytical method using Reverse Phase High Performance Liquid Chromatography coupled with Mass Spectrometry (RP-HPLC–MS/MS) for the simultaneous determination of Esomeprazole (ESOM) and Pantoprazole (PAN) in plasma. The method employs a quadrupole MS/MS system with electrospray ionization in positive mode to achieve highly sensitive and selective detection. Calibration curves were constructed over a concentration range of 0.5 to 2000 ppb for ESOM and PAN, with internal standard (IS) validation for improved accuracy. The method underwent thorough validation following ICH M10 guidelines, including specificity, linearity, accuracy, precision, and matrix effects. The results showed no significant interference from matrix components, confirming high specificity for both drugs. Precision and accuracy were consistently within acceptable limits, with %RSD values below 15% across all quality control levels. The lower limit of quantification (LLOQ) was determined to be 500 ppt for ESOM, ensuring the method's sensitivity for trace-level detection. Stability tests demonstrated that the analytes remained stable in plasma samples under various storage conditions. The method was successfully applied to clinical samples, providing reliable pharmacokinetic data. This RP-HPLC–MS/MS method is robust, precise, and sensitive, making it suitable for routine therapeutic drug monitoring and pharmacokinetic profiling of Esomeprazole and Pantoprazole in plasma.
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