Analytical Method Validation and Bioequivalence Analysis of Mesalazine in Human Plasma via LC-MS/MS
Keywords:
LC-MS/MS, mesalazine, bioequivalence, analytical method validation, Pharmacokinetics, plasma analysis, specificityAbstract
This study details the development, validation, and application of a robust analytical method for quantifying mesalazine in human plasma using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The method was optimized for specificity and sensitivity, achieving a lower limit of quantification (LLOQ) at 1 ppb, which complies with the International Council for Harmonisation (ICH M10) and European Medicines Evaluation Agency (EMEA) guidelines. Calibration curves were linear across a range of 1–160 ppb with a coefficient of determination (R²) exceeding 0.99. Analytical parameters, including precision, accuracy, carry-over, and matrix effects, were rigorously validated, demonstrating reliable performance. Short-term and long-term stability tests confirmed the analyte's robustness under varying conditions, including freeze-thaw cycles.
The method's clinical relevance was evaluated through a bioequivalence study involving 26 healthy volunteers, who received test and reference formulations of mesalazine. Plasma samples were collected at multiple intervals and analyzed to generate pharmacokinetic profiles, including key metrics such as maximum concentration (Cmax) and area under the curve (AUC). Results demonstrated comparable pharmacokinetic properties, confirming the bioequivalence of the test formulation with the reference product.
This validated method's high sensitivity and specificity make it suitable for both clinical and regulatory applications. The use of a stable internal standard, diazepam, mitigated potential analytical errors, ensuring precise quantification of mesalazine. This study not only underscores the method's robustness but also provides essential data supporting its utility in therapeutic monitoring, drug development, and regulatory compliance. The findings contribute significantly to advancing the field of bioanalysis and ensuring drug safety and efficacy through rigorous evaluation methods.
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