Quantitative Analysis of Residual EDTA in Vaccine Products by HPLC with Luminescence Detection of Terbium(III) Complexation

Abstract To facilitate the investigation of EDTA clearance in a protein purification process, a relatively simple, sensitive, and rapid method for EDTA analysis was developed using HPLC with pre‐column complexation of EDTA and terbium(III) (Tb3+). The limit of detection of this method is about 87 ng/mL, and the limit of quantitation is about 260 ng/mL. The accuracy, intra‐ and inter‐assay variability of this method were evaluated. The method was applied to determine the residual EDTA concentrations in various stages of the purification process for vaccine products. Examples of evaluating EDTA clearance in a protein purification process will be discussed.