Desarrollo y Validación de métodos espectrofotométricos UV/VIS para la cuantificación de Rifampicina, Isoniazida y Pirazinamida en estudios de bioexención
No Thumbnail Available
Date
2024
Authors
Journal Title
Journal ISSN
Volume Title
Publisher
Universidad Nacional de Trujillo
Abstract
La bioexención se caracteriza por la sustitución de estudios de bioequivalencia por pruebas
de disolución in vitro, basándose en el Sistema de Clasificación Biofarmacéutica. Estos
estudios necesitan métodos de cuantificación validados para dar confiabilidad a los
resultados. El objetivo de esta investigación fue desarrollar y validar métodos
espectrofotometricos UV/VIS para la cuantificación de Rifampicina, Isoniazida y
Pirazinamida para estudios de bioexención. La especificidad, linealidad, rango, exactitud,
precisión, estabilidad de la solución analítica, prueba de filtro, límites de detección y
cuantificación fueron evualuados según los estándares establecidos por los lineamientos de
la International Conference on Harmonisation y la Farmacopea de los Estados Unidos. Se
utilizaron diferentes condiciones de disolución y tiempos de muestreo. Se determinaron las
longitudes de onda de máxima absorbancia para los tres medicamentos en los tres medios de
disolución. El método fue lineal en los rangos de concentración estudiados. Los límites de
detección y cuantificación estuvieron dentro de lo esperado. Los valores recuperados se
encontraron entre 95%-105%. No se observó interferencia de los placebos. Los valores de
coeficientes de variación porcentual fueron menores al 2% para la precisión. Para la robustez
se evaluaron diferentes condiciones como diferentes longitudes de onda, cambios en la
concentración y los filtros Whatman No 40, No 41, No 42; los cuales no modificaron la
respuesta del método. Las muestras se conservaron sin cambios durante las 6 horas de trabajo
a 25 ºC. Sin embargo para rifampicina e isoniazida estas no fueron estables a 24 horas. Los
métodos fueron lineales, exactos, específicos y precisos en los rangos de concentración
estudiados y en los tres medios de disolución siendo apto su uso para estudios de
bioexención.
Biowaiver is characterized by the substitution of bioequivalence studies by in vitro dissolution tests, based on the Biopharmaceutical Classification System. These studies need validated quantification methods to give reliability to the results. The objective of this research was to develop and validate UV/VIS spectrophotometric methods for the quantification of Rifampicin, Isoniazid and Pyrazinamide for biowaiver studies. Specificity, linearity, range, accuracy, precision, stability of the analytical solution, filter test, detection and quantification limits were evaluated according to the standards established by the guidelines of the International Conference on Harmonisation and the United States Pharmacopeia. Different dissolution conditions and sampling times were used. The wavelengths of maximum absorbance were determined for the three drugs in the threedissolution media. The method was linear in the concentration ranges studied. The detection and quantification limits were within those expected. The recovered values were between 95%-105%. No interference from placebos was observed. The coefficients of variation percentage values were less than 2% for precision. For robustness, different conditions were evaluated such as different wavelengths, changes in concentration and Whatman filters No. 40, No. 41, No. 42; which did not modify the response of the method. The samples were preserved without changes during the 6 hours of work at 25 ºC. However, for rifampicin and isoniazid, these were not stable at 24 hours. The methods were linear, exact, specific and precise in the concentration ranges studied and in the three-dissolution media, being suitable for use in biowaiver studies.
Biowaiver is characterized by the substitution of bioequivalence studies by in vitro dissolution tests, based on the Biopharmaceutical Classification System. These studies need validated quantification methods to give reliability to the results. The objective of this research was to develop and validate UV/VIS spectrophotometric methods for the quantification of Rifampicin, Isoniazid and Pyrazinamide for biowaiver studies. Specificity, linearity, range, accuracy, precision, stability of the analytical solution, filter test, detection and quantification limits were evaluated according to the standards established by the guidelines of the International Conference on Harmonisation and the United States Pharmacopeia. Different dissolution conditions and sampling times were used. The wavelengths of maximum absorbance were determined for the three drugs in the threedissolution media. The method was linear in the concentration ranges studied. The detection and quantification limits were within those expected. The recovered values were between 95%-105%. No interference from placebos was observed. The coefficients of variation percentage values were less than 2% for precision. For robustness, different conditions were evaluated such as different wavelengths, changes in concentration and Whatman filters No. 40, No. 41, No. 42; which did not modify the response of the method. The samples were preserved without changes during the 6 hours of work at 25 ºC. However, for rifampicin and isoniazid, these were not stable at 24 hours. The methods were linear, exact, specific and precise in the concentration ranges studied and in the three-dissolution media, being suitable for use in biowaiver studies.
Description
Keywords
Medicamentos antituberculosos, Fármaco disuelto, Bioequivalencia, Solubilidad, Espectrofotometría