Analytical method validation of reversed phase HPLC for quantitative analysis of tartrazine and auramine o in powder drinks

Abstract

This study was aimed to perform analytical method validation of high-performance liquid chromatography (HPLC) technique using photo-diode array detector for the simultaneous determination of Tartrazine (TAR) and Auramine O (AUO) in powder drink products. TAR and AUO were analysed using Waters Shield C18 column (250 mm x 4.6 mm i.d., 5 µm) using PDA detector at 300-650 nm. The mobile phase used was acetonitrile-ammonium acetate 19 mM (86:14 v/v) delivered isocratically at a flow rate of 1.2 mL/ min. The optimized HPLC condition was subjected to analytical method validation by assessing some performance characteristics as guided by International Conference on Harmonization (ICH). The method was linear over the studied concentration ranges with the coefficient of determination (R2) of 0.999 and 0.997 for TAR and AUR with % intercept less than 2%, respectively. The developed method was sensitive with a limit of detection value of 0.0325 μg/mL and 0.1052 μg/mL for TAR and AUO, respectively. The method is also accurate and precise as indicated with acceptable recovery values of 99.0-100.7% for TAR and 102.1-106.5% for AUO with relative standard deviation (RSD) values lower than those required by Association of Official Analytical Chemists’ (AOAC). The developed method is simple and can be used for routine analysis of TAR and AUO for quality assurance purposes of powder drinks.