HPLC-UV Method Development for the Determination of EDTA in Oxycodone HCl Oral Liquids with Derivatization Technique. Robustness by Design of Experiments Approach

This study deals with a simple and precise analytical method development and validation of a comprehensive high performance liquid chromatography method for estimation of Edetate disodium (EDTA) in liquid formulations. The effective chromatographic separation of EDTA was achieved with a X Bridge C-8 column with isocratic elution of mobile phase composed of buffer, methanol and water (175:90:675 v/v/v). The buffer used in mobile phase contained 0.06 % of tetra butyl ammonium hydroxide and adjusted pH to 7.5 ± 0.05 with 86 % of ortho phosphoric acid. The HPLC instrument flow rate was 2.0 mL min−1 and UV-detector wavelength was set 259 nm for the quantification of EDTA. The column oven temperature was 45°C. The derivatization of EDTA was carried out using 1 % copper acetate solution to form a UV active chromophore group. Proved the robustness of the method by using Design of Experiments study. The correlation coefficient for EDTA was >0.9998 and linearity range was found to be 25-75 μg mL−1. The mean recovery values were found to 98.3 (50 % level), 98.8 (100 % level) and 98.4 (150 % level). Further, specificity of the current method was proved by separating the placebo peaks. The proposed validated HPLC method was successfully applied for the determination of EDTA in Oxycodon HCl oral solutions.