Validation of the high performance liquid chromatography method for the analysis of neomycin sulfate with resonance Rayleigh scattering detection

Abstract

A simple and rapid reversed-phase high performance liquid chromatography method, using resonance Rayleigh scattering detection, for the determination of neomycin sulfate in serum samples was developed. Separation was performed on a Synergi Hydro-RP column with water solution which contained 0.18% TFA (v/v) as the mobile phase. Separation variables were optimized and discussed. In acid medium, neomycin sulfate could react with trypan red to form ion-association complexes, there is a stoichiometric ratio of 1 : 1 between the reaction of the neomycin sulfate with trypan red by Job plot analysis and the molar ratio method. Scanning electron microscopy revealed that these aggregates were spherical gathered particles with an average size of 45 nm, which was in agreement with the theoretical results. The analytical parameters were provided by the system, limit of detection (LOD) of 56 ng mL⁻¹ (3σ), linear correlation of 0.9990 and linear range response from 0.48 to 23.52 μg mL⁻¹. The precision of the method was <4%. Herein we demonstrated a novel method for the determination of neomycin sulfate which obtained satisfactory results.