Achieving High‑Resolution Separation of Oligonucleotides with Novel PRP‑Z2 HPLC Columns
Oligonucleotide therapeutics and advanced nucleic acid tools have rapidly transitioned from niche research reagents to key components of commercial drug develop- ment pipelines. Consequently, contract development and manufacturing organizations (CDMOs) and institutional research cores face increasing demand for sophisticated chromatographic support, including high-resolution impurity profiling, length ladder characterization, and robust system suitability testing across a broad range of oligonucleotide sizes and chemical modifications.
These expanding requirements place significant stress on conventional reversed-phase LC platforms. Silica-based stationary phases, which dominate small- molecule separations, are inherently limited by pH and temperature instability. Furthermore, silica phases are prone to hydrolytic degradation and progressive loss of efficiency under typical oligonucleotide separation conditions (>60°C and ion-pairing agents). The exposure of residual silanols to the separation conditions, leaves active metal sites from degraded silica, which can introduce secondary interactions with the anionic phosphate backbone of oligonucleotides, leading to peak tailing, variable recoveries, and difficulties in quantifying low-level impurities.
