RESUMO
A simple, efficient, accurate and selective HPLC method has been developed and validated successfully for the estimation of 5-fluorouracil in rabbit plasma. The drug was eluted by using Supelco C18 column (1.5 cm x 4.6 mm, 5 µm) with a mobile phase consisting of methanol and water (20: 80, v/v) by adjusting pH at 3.2, using perchloric acid solution. The retention time was found to be 4.107 with a flow rate of I mL/min. Multiple validation parameters evaluated with high accuracy indicating that the current method sufficiently qualifies the recommended criteria. Regression value obtained from linearity curve was R² = 0.999 and percentage recovery obtained was ranged from 96.6 to 102.5%. A fine response in short run time with perfect resolution made the method highly suitable for pharmacokinetic studies.
Assuntos
Antimetabólitos Antineoplásicos/sangue , Cromatografia Líquida de Alta Pressão , Cromatografia de Fase Reversa , Monitoramento de Medicamentos/métodos , Fluoruracila/sangue , Animais , Antimetabólitos Antineoplásicos/farmacocinética , Soluções Tampão , Calibragem , Cromatografia Líquida de Alta Pressão/normas , Cromatografia de Fase Reversa/normas , Fluoruracila/farmacocinética , Concentração de Íons de Hidrogênio , Limite de Detecção , Modelos Lineares , Metanol/química , Percloratos/química , Coelhos , Padrões de Referência , Reprodutibilidade dos Testes , Solventes/química , Água/químicaRESUMO
The present study focused to develop rapid, accurate and sensitive reversed-phase high pressure liquid chromatography method for the quantification of esomeprazole (ESO) magnesium in rabbit plasma. Chromatographic separation was achieved isocratically on a reversed-phase C,, column using simple mobile phase consisting of of methanol : acetonitrile: 0.05 M phosphate buffer, pH 7 adjusted with potassium hydroxide (45 : 10 : 45, v/v/v) at a flow rate of 1.0 mL/min and UV detection at 302 nm. The method was validated for system suitability, linearity, precision, accuracy, stability, robustness, LOD and LOQ. The described method stated good linearity over the range of 0.01 to 2.5 pg/mL (r = 0.999). The extraction recovery of esomeprazole was more than 95.3%. The method was precise with relative standard deviation < 1% with more than 90% accuracy and limit of quantification 0.0309 µg/mL. The freeze thaw stability studies indicated that the rabbit plasma samples containing esomeprazole could be stored in freezer at -20°C and handled under normal laboratory conditions without significant loss of drug. In conclusion, the developed method is simple, cost effective and reproducible, with improved sensitivity and running time of analysis.