Method Development for the Characterization of Synthetic Oligonucleotides by LC-MS

September 13, 2020
Phenomenex

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In this presentation, we will demonstrate the impact of mobile phase optimization on chromatographic separation and MS sensitivity and spectral quality. Additionally, we will look into how temperature and flow-rate can impact oligo separations as well. Live: Thursday, October 1, 2020 at 11am EDT | 8am PDT | 4pm BST | 5pm CEST On demand available after final airing until Oct. 1, 2021

Register free: https://www.chromatographyonline.com/lcgc_w/oligonucleotides

Event Overview:

The primary method for quantitation and characterization of synthetic oligonucleotides is ion-pair reversed phase liquid chromatography (IP-RPLC). Mobile phase composition is typically an alkylamine such as triethylamine (TEA) and perfluorinated alcohol such as hexafluoroisopropanol (HFIP), with methanol as the strong solvent.

Often, method development for chromatographic separation of oligonucleotides consists primarily of adjustments to gradient program; i.e. shallow gradient slopes of 0.5-1 % B per column volume are utilized to obtain separation of closely related impurities. However, rarely are other method parameters explored to improved chromatographic and mass spectrometric performance.

In this presentation, we will demonstrate the impact of mobile phase optimization on chromatographic separation and MS sensitivity and spectral quality. Additionally, we will look into how temperature and flow-rate can impact oligo separations as well.

Key Learning Objectives:

  • Determine experimental parameters to ensure proper experimental design during method development
  • Understand the role of mobile phase and column chemistry in LC-MS methods for oligonucleotides
  • Learn how to improve overall method robustness and sensitivity for oligonucleotide method development

Speaker: Brian Rivera, Senior Product Manager- Biologics, Phenomenex

Time and Date: Live: Thursday, October 1, 2020 at 11am EDT | 8am PDT | 4pm BST | 5pm CEST

On demand available after final airing until Oct. 1, 2021

Sponsor: Phenomenex

Register free: https://www.chromatographyonline.com/lcgc_w/oligonucleotides

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