High-performance liquid chromatographic determination of deoxycytidine monophosphate and methyldeoxycytidine monophosphate for DNA demethylation monitoring: experimental design and artificial neural networks optimisation
Language English Country Netherlands Media print
Document type Comparative Study, Journal Article, Research Support, Non-U.S. Gov't
- MeSH
- Deoxycytidine analogs & derivatives chemistry MeSH
- Deoxycytidine Monophosphate chemistry MeSH
- DNA Methylation * MeSH
- Neural Networks, Computer MeSH
- Chromatography, High Pressure Liquid methods MeSH
- Research Design MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Comparative Study MeSH
- Names of Substances
- 5-methyldeoxycytidine MeSH Browser
- Deoxycytidine MeSH
- Deoxycytidine Monophosphate MeSH
The optimal conditions for the separation of 2'-deoxycytidine-5'-monophosphate and 5-methyl-2'-deoxycytidine-5'-moncphosphate in the matrix of other natural occurring nucleotides after digest of DNA were investigated. Using experimental design combined with artificial neural networks, efficient optimisation of the HPLC separation conditions was performed. The mobile phase composition was optimised on the basis of its three components (concentration of phosphate, content of methanol and pH). The best separation was obtained with a mobile phase containing 50 mM phosphate, pH 5.5 and 6% methanol. The final resolution achieved between 2'-deoxycytidine-5'-monophosphate and 5-methyl-2'-deoxycytidine-5'-monophosphate was equal to 2.78. Finally, the optimised system was successfully tested on the nucleotide mixture solution to determine the methylation state of 2'-deoxycytidine-5'-monophosphate in DNA in the search for FMR1 gene changes.
References provided by Crossref.org
