Analysis of Oligonucleotides Using a Single Quadrupole Mass Spectrometer with Hydrophilic Interaction Chromatography (HILIC)

User Benefits

- Synthetic oligonucleotides and impurities can be analyzed using HILIC. - Comprehensive characterization of synthetic oligonucleotides can be executed using LCMS-2050 single quadrupole mass spectrometer and LabSolutions Insight Biologics. - Purity calculations for target components can be performed by utilizing MS spectra simultaneously acquired with UV chromatogram.

Introduction

In recent years, oligonucleotide therapeutics have been rapidly developed and have attracted attention as a new drug discovery modality. Comprehensive detection and identification of impurities generated during the synthesis process and having different base lengths from the target product are required. Reverse-phase ion-pair chromatography is widely used for the separation of target compounds from impurities. Liquid chromatography-mass spectrometry (LC-MS) is commonly used as well since complete separation of the target product and all impurities seems almost impossible. However, the development of analytical methods without using ion-pair reagents is expected because ion-pair reagents such as triethylamine have safety concerns and issues related to flow pass contamination of the instrument. In this article, HILIC was focused on as the separation mode because it does not require ion-pair reagent and employs a mobile phase suitable for liquid chromatography-mass spectrometry then impurity analysis of synthetic oligonucleotides using HILIC with the LCMS-2050 high-performance liquid chromatography-mass spectrometry system and LabSolutions Insight Biologics was performed.

July 23, 2026 GMT