Rapid and accurate determination of deoxyribonucleoside monophosphates from DNA using micellar electrokinetic chromatography with a cationic surfactant additive
| Autor: | Inchul Yang, Nansook Hong, Sang-Ryoul Park, Lian Hua Shi, Ji Seon Jeong, Sook-Kyung Kim |
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| Rok vydání: | 2011 |
| Předmět: |
Chromatography
Chemistry Oligonucleotide Stacking Cationic polymerization Analytical chemistry Deoxyribonucleosides DNA Standard solution Biochemistry Micellar electrokinetic chromatography Analytical Chemistry Surface-Active Agents Capillary electrophoresis Pulmonary surfactant Cations Calibration A-DNA Chromatography Micellar Electrokinetic Capillary |
| Zdroj: | Analytical and Bioanalytical Chemistry. 400:2131-2140 |
| ISSN: | 1618-2650 1618-2642 |
| DOI: | 10.1007/s00216-011-4915-5 |
| Popis: | A micellar electrokinetic chromatography (MEKC) method for rapid and accurate determination of 2'-deoxyribonucleoside 5'-monophosphates (dNMPs), four structural elements of DNA, is described. MEKC separation at an optimized pH enabled complete separation of four dNMPs. The use of a cationic surfactant additive for MEKC led to the reversal of EOF, which enhanced the migration velocities of the negatively charged dNMPs. Under the optimized condition, full-baseline separation of the four dNMPs assuring accurate peak integration was obtained within 5 min. For the given separation condition, pH-mediated on-column sample stacking was optimized and applied to enhance sensitivity up to 6-fold. Analytical precision was improved by spiking iothalamate as an internal standard. The accuracy of dNMP quantitation was ensured with dNMP standard solutions determined by inductively coupled plasma-optical emission spectroscopy that measured phosphorous quantity. Performance of the proposed method was ultimately proven by accurate quantitation of a DNA oligonucleotide that was enzymatically hydrolyzed prior to dNMP analysis. The proposed MEKC method turned out to be a reliable analytical method for dNMPs that features high speed, high sensitivity, and high precision, and could be utilized for high-accuracy determination of the amount of DNA as well as the base composition of DNA. |
| Databáze: | OpenAIRE |
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