RESUMEN
A simple, sensitive, precise, accurate and robust high performance liquid chromatographic method has been developed for simultaneous estimation of saxagliptin (SAXA) and dapagliflozin (DAPA) in pharmaceutical formulation. Design of experiments (DoE) was applied for multivariate optimization of the experimental conditions of RP-HPLC method. Risk assessment was performed to identify the critical method parameters. Three independent factors; mobile phase composition, flow rate and column temperature were used to design mathematical models. Central composite design (CCD) was used to study the response surface methodology and to study in depth the effects of these independent factors. Desirability function was used to simultaneously optimize the retention time and resolution of SAXA and DAPA. The optimized and predicted data from contour diagram consisted of acetonitrile and ortho phosphoric acid (0.1%) in the ratio of 50:50 respectively, at a flow rate of 0.98 ml/min and column temperature 31.4 °C. Using these optimum conditions baseline separation of both drugs with good resolution and run time of less than 6 min were achieved. The optimized assay conditions were validated according to ICH guidelines. Hence, the results clearly showed that Quality by design approach could be successfully applied to optimize RP-HPLC method for simultaneous estimation of SAXA and DAPA.
Asunto(s)
Preparaciones Farmacéuticas/clasificación , Cromatografía Líquida de Alta Presión/métodos , Diabetes Mellitus Tipo 2/clasificación , Comprimidos/administración & dosificación , Formas de Dosificación , Optimización de Procesos/métodosRESUMEN
Tephrosia purpurea (L.) Pers., Fabaceae, is claimed to be of use in the control and treatment of a variety of epileptic disorders in Indian system of medicine. The present study plans to systematically evaluate T. purpurea and to verify this claim. Status epilepticus was induced in male albino rats of Wistar strain by administration of pilocarpine (30 mg/kg, i.p.) 24 h after lithium chloride (3 mEq/kg, i.p.). Different doses of the extract of T. purpurea were administered orally one hour before the injection of pilocarpine. The severity of status epilepticus was observed and recorded every 15 min till 90 min and thereafter every 30 min till 180 min, using the scoring system. The in vivo lipid peroxidation of rat brain tissue was measured. The in vitro NO free radical scavenging activity of plant extract was assessed. The interaction between plant extract and 2-diphenyl-2-picryl hydrazyl (DPPH) was also observed for in vitro free radical scavenging activity. The severity of status epilepticus was reduced with the administration of ethanolic extract of T. purpurea. Ethanolic extract of the plant exhibited both in vivo and in vitro antioxidant activity. The ethanolic extract of T. purpurea was found to be useful to control lithium-pilocarpine induced status epilepticus in albino rats of Wistar strain.
Tephrosia purpurea (L.) Pers., Fabaceae, é conhecida pelo seu uso no controle e tratamento de uma variedade de distúrbios epilépticos no sistema indiano de medicina. O presente estudo pretende avaliar de forma sistemática T. purpurea e verificar essa alegação. Status epilepticus foi induzido em ratos albinos machos da linhagem Wistar pela administração de pilocarpina (30 mg/kg, i.p.) 24 h após o cloreto de lítio (3 mEq/kg, i.p.). Diferentes doses do extrato de T. purpurea foram administrados por via oral uma hora antes da injeção de pilocarpina. A gravidade do status epilepticus foi observada e registrada a cada 15 min até 90 min e, posteriormente, a cada 30 min até 180 min, utilizando um sistema de pontuação. A peroxidação lipídica in vivo do tecido cerebral de ratos foi avaliada. A atividade captadora de radicais livres do extrato da planta foi avaliada in vitro. A interação entre o extrato da planta e 2-difenil-2-picril hidrazil (DPPH) também foi observada in vitro para atividade sequestradora de radicais livres. A gravidade do status epilepticus foi reduzida com a administração do extrato etanólico da T. purpurea. Extrato etanólico da planta apresentou, tanto in vivo quanto in vitro atividade antioxidante. O extrato etanólico da T. purpurea parece ser útil no controle de lítio de status epilepticus induzido pela pilocarpina em ratos albinos da linhagem Wistar.