Abstract
Improved separations of the protected oligonucleotides are reported using a polar bonded CN phase with gradient elution (methylene chloride-methanol mobile phase). This packing yielded high recoveries, and overcame some of the problems associated with the use of highly polar silica columns. In addition, the application of reversed-phase and ion-pair high-performance liquid chromatography is demonstrated for the oligonucleotides and for the nucleosides and mononucleotides, respectively. Ion-pair chromatography was used to accomplish the simultaneous separation of the ribo- and deoxyribonucleosides and their mononucleotides. A novel method for the determination of oligomer chain length is also reported, using diesterase cleavage and ion-pair high-performance liquid chromatography.
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