Separation of synthetic phosphorothioate oligodeoxynucleotides from their oxygenated (phosphodiester) defect species by strong-anion-exchange high-performance liquid chromatography

Abstract A method is described which separates synthetic deoxynucleotide phosphorothioates from their oxygenated (phosphodiester, “P = O”) defect species by strong-anion-exchange chromatography, using novel “soft-base” anionic eluents. The method enables the qualitative and quantitative assessment of P = O content in phosphorothioate DNA, and represents a rapid and sample-conserving alternative to 31 P-NMR.