ResourceLife Sciences
Mobile phase optimization for the analysis of synthetic oligonucleotides by ion-pair reversed phase liquid chromatography
22 May 2023Most oligos are manufactured by solid phase synthesis, where nucleotides are added in a step-wise phase, leading to the characterization of the commonly associated n-1 and n+1 impurities. However, even with ion-pair to facilitate the analytical separation, for full characterization of process and product-related impurities for synthetic oligonucleotides, extensive method development and mobile phase optimization are required. This application note from Phenomenex explores ion-pair reversed phase, an effective technique for the characterization and quantitation of synthetic oligonucleotides.
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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.