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Nucleic acid separations using superficially porous silica particles.

Elizabeth D Close1, Alison O Nwokeoji1, Dafydd Milton2

  • 1Department of Chemical and Biological Engineering, ChELSI Institute, University of Sheffield, Mappin Street, Sheffield S1 3JD, UK.

Journal of Chromatography. A
|March 8, 2016
PubMed
Summary

This study explores using superficially porous silica particles with ion pair reverse-phase liquid chromatography for nucleic acid analysis. Different pore sizes optimize separation for various nucleic acids, from small oligonucleotides to large DNA/RNA molecules.

Keywords:
Ion pair reverse-phase chromatographyNucleic acidsOligonucleotidesRNA

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Area of Science:

  • Analytical Chemistry
  • Biochemistry
  • Chromatography

Background:

  • Ion pair reverse-phase liquid chromatography is a standard method for separating nucleic acids.
  • Various stationary phases exist, but superficially porous particles offer unique advantages.

Purpose of the Study:

  • To evaluate superficially porous silica particles for nucleic acid separation using ion pair reverse-phase liquid chromatography.
  • To determine the impact of pore size on the resolution of diverse nucleic acid types.

Main Methods:

  • Utilized superficially porous silica particles with varying pore sizes (80Å, 150Å, 400Å).
  • Employed ion pair reverse-phase liquid chromatography for analyzing oligonucleotides, oligoribonucleotides, phosphorothioate oligonucleotides, dsDNA, and RNA.
  • Investigated separation of therapeutic oligonucleotide impurities.

Main Results:

  • 80Å pore sizes optimized small oligonucleotide separation, suitable for mass spectrometry.
  • 150Å pore sizes improved resolution for larger oligonucleotides (>19mers).
  • 400Å pore sizes yielded optimal resolution for large dsDNA/RNA molecules.

Conclusions:

  • Superficially porous silica particles are effective for nucleic acid separations via ion pair reverse-phase liquid chromatography.
  • Pore size is a critical parameter influencing resolution for different nucleic acid sizes.
  • This approach aids in analyzing therapeutic oligonucleotides and their impurities.