The analysis of impurities is of great importance during the development and production of pharmaceuticals and fi ne chemicals. This Application Note shows that multiple heart-cutting 2D-LC is ideally suited for method development for impurity analysis of pharmaceuticals and fi ne chemicals. Using the Agilent 1290 Infi nity 2D-LC Solution, every peak detected after the fi rst-dimension separation was heart-cut and transferred to a second-dimension separation with different selectivity. This approach enabled the discovery of possible coeluting compounds. In addition, the identity of impurities could be confi rmed by spiking the sample with the suspected impurity. Agilent 1290 Infinity II Multisampler