High-resolution sampling 2D-LC for pharmaceutical impurity analysis

In this application note, Agilent addresses the challenge of analyzing impurities in low concentrations relative to an active pharmaceutical ingredient (API) for quality control purposes. When the impurities have structural similarities to the API and there is a significant concentration difference, achieving chromatographic separation and detection becomes difficult.

To overcome this challenge, Agilent introduces the Agilent 1290 Infinity II 2D-LC Solution, which offers the flexibility to switch between comprehensive (LCxLC), multiple heart-cutting (MHC), or high-resolution sampling 2D-LC (HiRes 2D-LC) techniques. In this specific application note, HiRes 2D-LC is utilized to separate two closely eluting compounds, with one compound present at a very low concentration and concealed beneath the more highly concentrated compound peak. The analysis involves chlorodifluorobenzoic acids as standard substances and deamidated insulin as a representative real sample.