Agilent presents new method for oligonucleotide purification using HPLC/MS
Agilent Technologies has introduced a new, cost-effective preparative HPLC/MS method for the fast and selective purification of synthetic oligonucleotides. The method reduces the need for expensive reagents in scalable production.
Agilent Technologies has developed a new method for purifying synthetic oligonucleotides (ONs) using preparative High-Performance Liquid Chromatography coupled with Mass Spectrometry (HPLC/MS).
The method aims to accelerate and enhance the purification process for oligonucleotides, which are crucial in life science and diagnostics research. Previous techniques, such as ion-pair reversed-phase HPLC (IP-RP HPLC), have been effective, but scaling them up for production required large volumes of expensive reagents like hexafluoroisopropanol (HFIP), limiting their cost-effectiveness.
Agilent's novel approach utilizes dibutylamine (DBA) as an ion-pairing agent and tris(hydroxymethyl)aminomethane (TRIS) as a buffer. DBA has demonstrated higher resolution with short-chain ONs, and TRIS replaces the costly HFIP. This substitution allows for the purification of larger batches at a reduced expense.
The process employs the Agilent 1290 Infinity II Autoscale Preparative LC/MSD system. Purification is achieved through automated software that optimizes gradients and collects fractions based on mass spectrometry data. Re-analysis of collected fractions confirms that the purified oligonucleotides typically reach a purity level exceeding 99%.
The company presented these findings in a poster detailing the method's capability for rapid and selective production of high-quality oligonucleotides, facilitating their broader application in research and diagnostics.