High Performance Liquid Chromatographic determination of deoxycytidine monophosphate and methyl-deoxycytidine monophosphate for DNA demethylation monitoring: Experimental Design and Artificial Neural Networks optimisation

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14310%2F01%3A00004613" target="_blank" >RIV/00216224:14310/01:00004613 - isvavai.cz</a>

Výsledek na webu

—

DOI - Digital Object Identifier

—

Jazyk výsledku

angličtina

Název v původním jazyce

High Performance Liquid Chromatographic determination of deoxycytidine monophosphate and methyl-deoxycytidine monophosphate for DNA demethylation monitoring: Experimental Design and Artificial Neural Networks optimisation

Popis výsledku v původním jazyce

The optimal conditions for the separation of 2'-deoxycytidine-5'-monophosphate and 5-methyl-2'-deoxycytidine-5'-monophosphate 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 base 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.

Název v anglickém jazyce

High Performance Liquid Chromatographic determination of deoxycytidine monophosphate and methyl-deoxycytidine monophosphate for DNA demethylation monitoring: Experimental Design and Artificial Neural Networks optimisation

Popis výsledku anglicky

The optimal conditions for the separation of 2'-deoxycytidine-5'-monophosphate and 5-methyl-2'-deoxycytidine-5'-monophosphate 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 base 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.

Druh

J_x - Nezařazeno - Článek v odborném periodiku (Jimp, Jsc a Jost)

CEP obor

CB - Analytická chemie, separace

OECD FORD obor

—

Projekt

—

Návaznosti

—

Rok uplatnění

2001

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

J. Chromatogr. B.

ISSN

0378-4347

e-ISSN

—

Svazek periodika

2001

Číslo periodika v rámci svazku

755

Stát vydavatele periodika

NL - Nizozemsko

Počet stran výsledku

9

Strana od-do

185

Kód UT WoS článku

—

EID výsledku v databázi Scopus

—