Immobilization of Adenosines on Silica Gel and Specific Separation of Thymidine Oligomers

  • Yoshiaki Inaki
  • Kenji Matsukawa
  • Kiichi Takemoto

Abstract

Adenosine and deoxyadenosine derivatives were immobilized on silica gel, which was able to be used as HPLC resins for the selective separation of oligothymidine, pd(T)n, from the mixture of oligonucleotides. The chromatograms were obtained in 10% methanol-phosphate buffer aqueous mobile phase through 5°C to 30°C. The longest retention time was observed for the complementary pd(T)4, and increased with decrease of temperature. The most hydrophobic pd(A)4 did not show a long retention time and showed a small temperature dependency. This fact suggested that the hydrophobic interaction in this system was negligible, and the main separation factor was the base pairing between complementary nucleic acid bases. The retention times of pd(T)n became longer with a decrease of temperature. The −ΔH value for pd(T)4 was 16.8 kcal/mol, which was approximately 4 kcal per one thymine base. An increase of the degree of polymerization caused retardation of the retention times, and gave higher −ΔH values. Interestingly, an increase of one thymidine unit {pd(T)2 to pd(T)3, and pd(T)3 to pd(T)4} caused an increasing −ΔH value of approximately 4 kcal, which is the same value obtained for pd(T)4. These resins may be useful for separation of components of nucleic acids and polynucleotides as a specific separation system.

Keywords

Longe Retention Time Acetic Acid Aqueous Solution Hoff Plot Specific Separation Thymine Base 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer Science+Business Media New York 1994

Authors and Affiliations

  • Yoshiaki Inaki
    • 1
  • Kenji Matsukawa
    • 1
  • Kiichi Takemoto
    • 1
  1. 1.Department of Applied Fine Chemistry Faculty of EngineeringOsaka UniversitySuita, Osaka 565Japan

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