Study of the retention behavior of trinucleotides in HPLC
| Authors | |
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| Year of publication | 2014 |
| Type | Article in Proceedings |
| Conference | Acta Universitatis Palackianae Olomucensis Chemica 51S |
| MU Faculty or unit | |
| Citation | |
| Field | Analytic chemistry |
| Keywords | trinucleotides; RP-HPLC |
| Description | Trinucleotides are short DNA sequences of three nucleobases; in our study we used guanine, adenine and thymine. It was shown that a useful tool to study the effect of sequence on retention was achieved by reversed phase liquid chromatography in ion-interaction mode (II-RPLC). Retention mechanism in the II-RPLC is based on addition of the ion interaction agent (in a form of buffer ions) to the mobile phase, which can effectively compensate the charge on the phosphate groups of a sugar-phosphate backbone, thereby increasing the hydrophobicity and thus the retention in system with reversed phase. To study the retention behavior we have chosen a combination of experimental design (ED) and artificial neural networks (ANN). ED was used to target more variable parameters in parallel while monitoring the system response. This method allows to determine the size of the effect of the various factors involved in the retention and to possibly reveal their mutual correlation. It is commonly connected to approximation of the response function using artificial neural networks (ANN), which then enables prediction of the retention time of an unknown sequence. This approach could be used as a tool to identify unknown short nucleotide sequences in mixtures |
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