Denaturing reversed-phase HPLC using a mobile phase containing urea for oligodeoxynucleotide analysis

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Denaturing reversed-phase (RP) high performance liquid chromatography (HPLC) is usually achieved by elevating column temperature. In this article, an alternative method involving using a mobile phase that contains urea and performing HPLC at room temperature is described. The efficacy of the new method was demonstrated by analyzing a 61-mer oligodeoxynucleotide (ODN) and double-stranded (ds) ODNs. The multiple peaks of the 61-mer ODN under normal conditions merged into one under the denaturing conditions. The broad single peaks of dsODNs under normal conditions were split into two sharp peaks.

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Copyright Taylor and Francis Group, LLC. Publisher’s version of record: https://doi.org/10.1080/15257770.2014.891740

Publication Title

Nucleosides, Nucleotides and Nucleic Acids