Column wall modifications using polyethylene oxide for capillary electrophoresis of ribonucleotides |
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Authors: | X Shao Y Shen K O'Neill M L Lee |
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Institution: | (1) Department of Microbiology, Brigham Young University, 84602 Provo, UT, USA;(2) Department of Chemistry and Biochemistry, Brigham Young University, 84602 Provo, UT, USA |
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Abstract: | Summary In this study, a variety of fused silica capillaries with different combinations and sequences of treatments with HMDS and
polyethylene oxide were prepared in order to develop an optimized column modification method for analysis of ribonucleotides.
The 12 most common ribonucleotides (UTP, CTP, ATP, GTP, UDP, CDP, ADP, GDP, UMP, CMP, AMP, and GMP) in human cells were used
as test solutes. Column performance measurements, including electroosmotic flow (EOF), solute migration speed and retention,
column efficiency, peak shape, and resolution were investigated. By analyzing solute migration speed and retention of various
hydrophilic/hydrophobic solutes, the column wall effects (EOF and adsorption) can be distinguished. This analysis method can
give guidance in optimizing polymer coating properties (hydrophilicity/hydrophobicity) for CE columns. By studying the performance
of these columns after various surface treatments, we were able to improve the separation of ribonucleotides from real samples
to within 16 minutes with high efficiency and stability (over 300 analyses) using columns first deactivated with hexamethyldisilazane,
and then coated with polyethylene oxide. |
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Keywords: | Capillary electrophoresis Column technology Deactivation Polyethyleneoxides Ribonucleotides |
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