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Publication Date

Summer 2010

Degree Type

Thesis - Campus Access Only

Degree Name

Master of Science (MS)

Department

Chemistry

Advisor

Joseph J. Pesek

Keywords

HPLC, Nucleotide separation, UDA column

Subject Areas

Chemistry, Analytical

Abstract

The UDA column, which has undecynoic acid groups attached to the silica-hydride surface, was chromatographically characterized in this research, and the retention mechanisms are discussed. The polar molecules tested on the UDA column were metabolites (amino acids, carbohydrates), peptides, proteins, nucleosides, nucleotides, and galactosemia biomarkers. Different instrumentation and retention modes were employed depending on the spectroscopic and structural properties of the solutes.

The interactions between the stationary phase and the solutes include both hydrophobic and ionic/hydrophilic interactions. The UDA column possesses a typical aqueous normal phase behavior that can retain both polar and non-polar compounds with a change of mobile-phase composition. The UDA column showed significant retention and separation of most of the hydrophilic compounds, especially nucleotides. Therefore, the UDA stationary phase is classified as an ideal stationary phase for nucleotide separation. However, for other types of compounds, such as a mixture of galactosemia biomarkers, the UDA column was found to be ineffective under the conditions used in this research.

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