High performance liquid chromatographic separation of oligonucleotides using deoxyadenosine immobilized silica gel for stationary phase

Yoshiaki Inaki, Kenji Matsukawa, Hong Wang, Eiko Mochizuki, Takehiko Wada, Kiichi Takemoto

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Deoxyadenosine immobilized silica gels were prepared as the novel HPLC packing materials for the separation of nucleic acid fragments. Tetrathymidylic acid 5'-monophosphate {pd(T)4} was clearly separated from the other tetradeoxynucleotides by the specific interaction of complementary nucleic acid bases. The separation factor (α) of pd(T)4 increased with the decrease of columun temperature from 30°C to 5°C, because the specific bondings between the immobilized ligands and the complementary solutes are stabilized with the decrease of temperature. The presence of the hydrophilic spacer group between stationary phase and the nucleoside ligand caused higher separation efficiency for the complementary solute, compared with the stationary phase having no hydrophilic spacer groups. The "trityl-on" deoxyadenosine as the ligand was preferred to the unprotected one, which may be related to the conformation of the adenosine on silica gel.

Original languageEnglish
Pages (from-to)99-105
Number of pages7
JournalBUNSEKI KAGAKU
Volume42
Issue number2
DOIs
Publication statusPublished - 1993 Jan 1
Externally publishedYes

Keywords

  • HPLC
  • adenosine-immobilized silica
  • hydrogen bonding
  • specific interaction
  • tetradeoxynucleotide

ASJC Scopus subject areas

  • Analytical Chemistry

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