RESUMEN
The development and validation of a column high-performance liquid chromatographic assay method for the determination of aspirin and clopidogrel in tablet formulation are described. The combination formulation was subjected to International Conference on Harmonization-recommended stress conditions. Separation of the drugs from the degradation products formed under stress conditions was achieved on an octasilyl (C8) column using 0.3% orthophosphoric acid-acetonitrile (65 + 35, v/v) mobile phase. The method was validated for specificity, linearity, limits of detection and quantification, precision, accuracy, and robustness. The method was found to be specific against placebo interference and during the forced degradation. The response was linear in the concentration range of 30.0-120.0 microg/mL for aspirin and 15.0-60.0 microg/mL for clopidogrel, with a correlation coefficient of 0.9999 for both. The relative standard deviation values for intra- and interday precision were <2.0%. The accuracy was between 99.12 and 99.83% for aspirin and 98.20 and 100.35% for clopidogrel. Stress testing showed degradation products that were well-separated from the parent compound, confirming the stability-indicating capacity of the method.
Asunto(s)
Aspirina/análisis , Cromatografía Líquida de Alta Presión/métodos , Comprimidos/análisis , Comprimidos/química , Ticlopidina/análogos & derivados , Aspirina/aislamiento & purificación , Clopidogrel , Concentración de Iones de Hidrógeno , Inhibidores de Agregación Plaquetaria/análisis , Inhibidores de Agregación Plaquetaria/aislamiento & purificación , Reproducibilidad de los Resultados , Estrés Mecánico , Ticlopidina/análisis , Ticlopidina/aislamiento & purificaciónRESUMEN
A simple, precise, and accurate isocratic reversed-phase (RP) stability-indicating column high-performance liquid chromatographic (HPLC) assay method was developed and validated for determination of nebivolol in solid pharmaceutical dosage forms. Isocratic RP-HPLC separation was achieved on a Phenomenex Luna C8 (2) column (250 mm x 4.6 mm id, 5 microm particle size) using mobile phase composed of acetonitrile-pH 3.5 phosphate buffer (35 + 65, v/v) at a flow rate of 1.0 mL/min, and detection was performed at 280 nm using a photodiode array detector. The drug was subjected to oxidation, hydrolysis, photolysis, and heat to apply stress conditions. The method was validated for specificity, linearity, precision, accuracy, robustness, and solution stability. The method was linear in the drug concentration range of 40-160 microg/mL with a correlation coefficient of 0.9999. The repeatability relative standard deviation (RSD) for 6 samples was 0.69%, and the intermediate precision (RSD) for 6 samples was 1.39%. The accuracy (recovery) was between 98.57 and 99.55%. Degradation products produced as a result of stress studies did not interfere with detection of nebivolol, and the assay can thus be considered stability-indicating.