Ceftaroline fosamil: development a rapid HPLC method indicating stability and bioassay for determination in pharmaceutical formulation, stability and cytotoxicity studies

Abstract

The present study reports the development and validation of a microbiological assay. To assess this methodology, the method was developed and validated for the quantification of CEF by high performance liquid chromatography (HPLC). The validation of the microbial assay by diffusion method in 3x3 cylinder agar presented showed satisfactory results as to specificity, linearity in the range of 2.0 - 8.0 μg.mL^-1, precision (109.42 %), accuracy (102.3 %), and robustness. The development and validation of the method by HPLC were evaluated according to specificity, linearity, precision, accuracy and robustness. A high performance liquid chromatography from Shimadzu with Agilent^® C18 column, mobile phase (water with triethylamine 1.0 % pH 5.0: acetonitrile 87:13 v/v was used in the chromatographic method. The validated microbiological and chromatographic methods were compared statistically and there was no significant difference between them when compared by Student's t-test. In the preliminary stability study, it was found stable in acid hydrolysis (0.1M) and UVA light in the period evaluated, and unstable against thermal degradation (40 and 60 °C), oxidative with hydrogen peroxide, basic in NaOH (0.1 M and 0.01M) and UVC light. Samples exposed to UVC light and thermal degradation at 60°C showed degradation kinetics following zero order and second order, respectively. The cytotoxicity assay showed no difference between the normal condition and the sample submitted to forced degradation, suggesting that the possible degradation products formed did not change the result. The methods developed did not present a significant difference, therefore, they are interchangeable, and so can be used for routine quality control analysis.