Mitigation of Non-Specific Adsorption During Column Conditioning for an Oligonucleotide HILIC Analysis

Oligonucleotides, the short strands of nucleic acids that are pivotal in genetic research and therapeutic applications, require precise analytical techniques for their characterization. Hydrophilic Interaction Liquid Chromatography (HILIC) has emerged as a powerful alternative to traditional ion-pair reversed phase chromatography for oligonucleotides analysis (<35 mer). HILIC offers several advantages, including reduced mobile phase cost, lower toxicity, and improved stability, making it a cost-effective and environmentally friendly option. However, oligonucleotides are known to interact with metal surfaces, which can lead to non-specific adsorption onto both the column and LC hardware and negatively impact recovery and peak shape. Historically, one solution to non-specific adsorption was passivation, which is the process of masking active sites of metallic surfaces, but this is time-consuming and not longstanding. Another solution to address non-specific adsorption is the use of MaxPeak High Performance Surfaces (HPS) Technology, which offers improvements in separation and detection of metal-sensitive analytes. This application note focuses on the analysis of oligonucleotides using HILIC conditions to compare traditional stainless-steel High Performance Liquid Chromatography (HPLC) Systems and columns with systems and columns employing MaxPeak HPS Technology.

Benefits
- Increased recovery of oligonucleotides with “out-of-the-box" performance using MaxPeak HPS Technology on both system and column without the need for passivation or ion-pairing reagents
- HILIC separation of oligonucleotides uses fewer toxic solvents, contributing to a greener laboratory environment while also reducing operational costs
- Improved robustness and stability for HILIC separation of oligonucleotides with the Alliance™ iS Bio HPLC System and the MaxPeak HPS Column