Developing an improved UHPLC method for impurity profile analysis of ceftriaxone using analytical quality by design.

Abstract

In this study, we developed a new reversed-phase ultra-high-performance liquid chromatography (UHPLC) method for comprehensively measuring impurities in ceftriaxone. The method was developed based on the Chinese Pharmacopoeia (ChP) HPLC method, which is limited by the lack of selectivity to potential impurities and a long running time. Screening experiments showed that octylamine concentration, mobile phase pH, and organic phase ratio were critical method parameters. Further optimisation and Monte-Carlo simulations were performed to map out the design space. The selected working conditions resulted in a complete separation of the impurity profile in approximately 10 min. A multivariate approach confirmed that the method was robust, and the proportion of acetonitrile should be carefully controlled. Additionally, the developed UHPLC method could be transferred back to HPLC in a single step using a Columns Calculator, providing a new approach for the rapid and effective development of the HPLC method. Our findings could serve as a reference for developing the next version of the ChP.