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Emerging alternative analytical techniques for oligonucleotide separation and purification

Agilent Technologies
29 Apr 2025

As the therapeutic oligonucleotide market continues to mature, the need for alternative analytical approaches emerge to meet those demands. The current standard widely used for oligonucleotide analysis is ion-pairing reversed-phase chromatography (IP-RP) and ion-exchange chromatography (IEX). However, these instruments come with limitations such as, incompatibility and flexibility.

Explore an effective and emerging technique for oligonucleotide workflows – hydrophilic interaction chromatography (HILIC) and size exclusion chromatography (SEC) – and uncover the benefits it can provide for your separation and purification of oligonucleotides.

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Poroshell 120

Agilent Technologies

Poroshell 120 columns provide rugged, high resolution separations within the pressure range of any mainstream LC system, making the benefits of sub-2 µm performance available with all existing LCs.Poroshell columns can achieve high resolution and high speed separations on current instruments, and higher resolution and speeds on new high-pressure LC and LC/MS systems. Stable to 600 bar, Poroshell 120 are great for fast UHPLC analyses, and can accelerate your older, traditional 5 µm method for 4-5x reduction in analysis time. With a family of ten chemistries, including the recently introduced PFP, and alignment with the ZORBAX family chemistries, method development and transfer is flexible and easy.Features: High efficiency and high resolution, with up to 50% less backpressure than sub-2 μm columns 2 μm frit, for rugged performance with dirty samples Compatible with 400 bar and 600 bar LCs, as well as UHPLC instruments An expanding family of bonded phases to align with the ZORBAX Family, for reliable scalability Excellent selectivity and peak shapes Designed for exceptional reproducibility Explore Poroshell 120 solutions focused applications, from food to pharma

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OligonucleotidesOligonucleotides are small nucleic acid polymers, usually less than 20 bases in length. Oligonucleotides can be made via enzymatic cleavage or more commonly by chemical synthesis with polymerases. Their use includes FISH, southern blots, microarrays and as primers in PCR. High fidelity synthesis kits and detection systems are available for easy production and detection, respectively.Size Exclusion ChromatographySize exclusion chromatography (SEC) is a liquid chromatographic technique used to separate and identify molecules based on their size, including polymers, proteins, nanoparticles & nucleic acid. SEC is further subdivided into gel filtration chromatography (GFC) and gel permeation chromatography (GPC) when an aqueous or organic mobile phase is used respectively. HPLC-based SEC systems and columns are available for analytical separation alongside SEC spin columns or filters to prep samples for many protocols. Find the best size exclusion chromatography equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Mass SpectrometryMass spectrometry (MS) is a powerful analytical technique used to identify and quantify molecules based on the mass-to-charge ratio of gas-phase ions. It provides detailed information about the structure, composition, and properties of compounds and is widely used across fields such as environmental monitoring, materials science, drug discovery and development, food and beverage testing, and wider chemical research. Key MS techniques include tandem mass spectrometry (MS/MS), liquid chromatography–mass spectrometry (LS-MS) and inductively coupled plasma (ICP-MS). Choosing from these wide range of techniques and technologies can be a daunting task, so keep up to date with scientific applications, performance expectations, and customer reviews here all in one place. Visit our product directory to receive quotes direct from the manufacturer. bio-separation analysesBiopharmaceuticalsBiopharmaceuticals are proteins and other compounds (such as nucleic acids) produced by living organisms that have uses as therapeutics or for in vivo diagnostics. The most well known example of a biopharmaceutical product, and the first to be approved for therapeutic use, was recombinant human insulin.

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Emerging alternative analytical techniques for oligonucleotide separation and purification