Poster Presentations
- Day 3, May 13（Wed.） Poster (1008/09)
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3P-37 PDF
Examination of Analytical Conditions for Separation of Oligonucleotides by Hydrophilic Interaction Chromatography coupled to Mass Spectrometry
For the quality control of synthetic oligonucleotides, ion-pairing reversed phase chromatography (IP-RPC) and ion exchange chromatography (IEC) are commonly used. IP-RPC using triethylamine and hexafluoroisopropanol (HFIP) as additives in the eluents is usually used for mass spectrometry detection. However, under this RPC conditions, the separation selectivity of homologues or isomers formed by phosphorothioation is lower than that of IEC. For improvement of the separation selectivity of analytes under IP-RPC, the method using trifluoroethanol (TFE) as an alternative to HFIP was reported. In this study, we examined hydrophilic interaction chromatography (HILIC) as another approach. The separation of analytes was performed using HILIC column packed with hydrophilic stationary phase of silica-gel with carbamoyl group. As eluents, ammonium formate solution and acetonitrile were used. As a result of examining the optimum analytical conditions, the separation selectivity of homologues or isomers formed by phosphorothioation was almost equivalent to that of the IP-RPC (using TFE) or IEC methods. It was suggested that HILC conditions may be useful for oligonucleotide analysis.