Krisztián Horváth

High performance liquid chromatography (HPLC) is a widely used and well-established technique, routinely employed by thousands of analytical scientists worldwide. Nonetheless, certain challenges—arising from the complex interplay of multiple factors affecting peak retention and separation—persist. In this context, multidimensional modeling approaches can provide valuable support.

Serial coupling of different column types can provide several important benefits to the chromatographer, including fine-tuning selectivity to separate complex mixtures.

The impact of ionic strength, buffer capacity, and pH-response on the retention behavior and peak shape of mAb species characterization is evaluated for IEX-MS. The aim of the present study was to understand the impact of ionic strength, buffer capacity, and pH-response on the retention behavior and peak shape of mAb species.

Characterization of mAbs and related products requires the identification of chromatographic peaks with MS. However, the conventional salt- and pH-gradient elution techniques used in IEX are inherently incompatible with MS. Ammonium acetate- and ammonium carbonate-based mobile phase systems have been recently applied in IEX-MS, but the influence of the eluent composition on peak shape and retention has not been discussed nor studied systematically until now. The aim of the present study was to understand the impact of ionic strength, buffer capacity, and pH-response on the retention behaviour and peak shape of mAb species.