Mobile phase optimization for the analysis of synthetic oligonucleotides by ion-pair reversed phase liquid chromatography

Most oligos are manufactured by solid phase synthesis, where nucleotides are added in a step-wise phase, leading to the characterization of the commonly associated n-1 and n+1 impurities. However, even with ion-pair to facilitate the analytical separation, for full characterization of process and product-related impurities for synthetic oligonucleotides, extensive method development and mobile phase optimization are required. This application note from Phenomenex explores ion-pair reversed phase, an effective technique for the characterization and quantitation of synthetic oligonucleotides.