Analytical Method Development and Validation for the Identification of Spiraeoside Using RP-HPLC in Pharmaceutical Gel Formulation

摘要

The aim of this study was to validate a Reversed-Phase High Performance Liquid Chromatographic(RP-HPLC)method for the qualitative determination of spiraeoside contained in a gel preparation.The chromatography column used was a stainless steel column(25 cm x 4.6 mm,dp=5 μm)packed with silica surface covered with cross-linked diol groups for polar selectivity(Luna HILIC).Mobile phase consisting buffer containing 1 mL of 10% Tetra methyl ammonium hydroxide aqueous solution(pH 2.5)and acetonitrile and isocratic system was used.The flow rate was 0.4 ml/min and the detective wavelength was 365 nm.Injection value for each time was 100 μl and separations were carried out isothermally at 35℃ in a heated chamber and sample temperature was 25℃.System suitability parameters were studied by injecting the standard solution six times and results were well under the acceptance criteria.The proposed validated method could separate spiraeoside peak was confirmed using photo-diode array detector.