Efficient Method Development of Oligonucleotides by Reversed-Phase Ion-Pair Chromatography

• LabSolutions MD can improve the efficiency of method development for oligonucleotides and related impurities.
• LCMS-2050 single quadrupole mass spectrometer can accurately track each peak of oligonucleotides and related impurities.
• Nexera™ XS inert (UHPLC system) with Shim-pack Scepter™ Claris (inert-coating metal-free column) offers complete inertness of the sample flow path to achieve optimal chromatographic separation of oligonucleotides

Nucleic acid drugs, such as antisense oligonucleotides, exert their effect by interacting with targets (genes and proteins) inside and outside of cells. Nucleic acid drugs are produced through chemical synthesis, but the synthesis process can introduce impurities such as shorter and longer length of products and protection groups. Therefore, proper separation of the target oligonucleotide is required.

For LC separation, one commonly used mode is reversed-phase ion-pair chromatography (RP-IP). The separation patterns obtained with RP-IP chromatography can vary depending on the concentration of the ion pair reagent and the composition of organic solvent. In addition, the separation behavior can differ based on the length of products, nucleobase, and the presence of modifications. Therefore, it is important to optimize the separation for each sequence of oligonucleotides. This article describes how to achieve the optimal separation of oligonucleotides and related impurities efficiently by utilizing LabSolutions MD, a dedicated software for supporting method development, through initial screening and optimization phase respectively.

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