Advanced search
1 file | 286.83 KB

Separation of purine and pyrimidine bases by ion chromatography with direct conductivity detection

(2008) JOURNAL OF CHROMATOGRAPHY A. 1193(1-2). p.104-108
Author
Organization
Abstract
A simple, rapid and accurate method for the simultaneous determination of four purine and pyrimidine bases (cytosine, 5-methylcytosine, adenine and N6-methyladenine) has been developed. The quantitative determination of these bases was accomplished by ion chromatography (IC) with direct conductivity detection (CD) based on their ionization in acidic medium without chemical suppression. The recovery of cytosine, 5-methylcytosine, and adenine in calf thymus DNA was more than 98% (n = 3) and the relative standard deviation (RSD, n = 5) less than 2.4%. In a single chromatographic run, the four bases could be separated and determined in less than 10 min. The detection limits were found to be 0.05 mu g/mL for cytosine, 0.08 mu g/mL for 5-methylcytosine, 0.07 mu g/mL for adenine, and 0.07 mu g/mL for N6-methyladenine. Linear ranges were 0.2-95.1 mu g/mL for cytosine (r(2) = 0.9996),0.3-196.6 mu g/mL for 5-methylcytosine (r(2) = 0.9994), 0.3-105.5 mu g/mL for adenine (r(2) = 0.9998), and 0.3-159.1 mu g/mL for N6-methyladenine (r(2) = 0.9999). With the proposed method, purine and pyrimidine bases could be successfully detected in calf thymus DNA. We also determined these bases in calf thymus DNA using RP-HPLC. Compared to RP-HPLC, the IC method offers advantages such as high selectivity and simple mobile phase.
Keywords
5-methylcytosine, cytosine, adenine, N6-methyladenine, ion chromatography, conductivity detection, PERFORMANCE LIQUID-CHROMATOGRAPHY, DNA METHYLATION, CANCER, NUCLEOSIDES, CORDYCEPS, SILICA

Downloads

  • (...).pdf
    • full text
    • |
    • UGent only
    • |
    • PDF
    • |
    • 286.83 KB

Citation

Please use this url to cite or link to this publication:

Chicago
Liu, L, J Ouyang, and Willy Baeyens. 2008. “Separation of Purine and Pyrimidine Bases by Ion Chromatography with Direct Conductivity Detection.” Journal of Chromatography A 1193 (1-2): 104–108.
APA
Liu, L, Ouyang, J., & Baeyens, W. (2008). Separation of purine and pyrimidine bases by ion chromatography with direct conductivity detection. JOURNAL OF CHROMATOGRAPHY A, 1193(1-2), 104–108.
Vancouver
1.
Liu L, Ouyang J, Baeyens W. Separation of purine and pyrimidine bases by ion chromatography with direct conductivity detection. JOURNAL OF CHROMATOGRAPHY A. PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS: Elsevier Science; 2008;1193(1-2):104–8.
MLA
Liu, L, J Ouyang, and Willy Baeyens. “Separation of Purine and Pyrimidine Bases by Ion Chromatography with Direct Conductivity Detection.” JOURNAL OF CHROMATOGRAPHY A 1193.1-2 (2008): 104–108. Print.
@article{539305,
  abstract     = {A simple, rapid and accurate method for the simultaneous determination of four purine and pyrimidine bases (cytosine, 5-methylcytosine, adenine and N6-methyladenine) has been developed. The quantitative determination of these bases was accomplished by ion chromatography (IC) with direct conductivity detection (CD) based on their ionization in acidic medium without chemical suppression. The recovery of cytosine, 5-methylcytosine, and adenine in calf thymus DNA was more than 98\% (n = 3) and the relative standard deviation (RSD, n = 5) less than 2.4\%. In a single chromatographic run, the four bases could be separated and determined in less than 10 min. The detection limits were found to be 0.05 mu g/mL for cytosine, 0.08 mu g/mL for 5-methylcytosine, 0.07 mu g/mL for adenine, and 0.07 mu g/mL for N6-methyladenine. Linear ranges were 0.2-95.1 mu g/mL for cytosine (r(2) = 0.9996),0.3-196.6 mu g/mL for 5-methylcytosine (r(2) = 0.9994), 0.3-105.5 mu g/mL for adenine (r(2) = 0.9998), and 0.3-159.1 mu g/mL for N6-methyladenine (r(2) = 0.9999). With the proposed method, purine and pyrimidine bases could be successfully detected in calf thymus DNA. We also determined these bases in calf thymus DNA using RP-HPLC. Compared to RP-HPLC, the IC method offers advantages such as high selectivity and simple mobile phase.},
  author       = {Liu, L and Ouyang, J and Baeyens, Willy},
  issn         = {0021-9673},
  journal      = {JOURNAL OF CHROMATOGRAPHY A},
  keyword      = {5-methylcytosine,cytosine,adenine,N6-methyladenine,ion chromatography,conductivity detection,PERFORMANCE LIQUID-CHROMATOGRAPHY,DNA METHYLATION,CANCER,NUCLEOSIDES,CORDYCEPS,SILICA},
  language     = {eng},
  number       = {1-2},
  pages        = {104--108},
  publisher    = {Elsevier Science},
  title        = {Separation of purine and pyrimidine bases by ion chromatography with direct conductivity detection},
  url          = {http://dx.doi.org/10.1016/j.chroma.2008.04.011},
  volume       = {1193},
  year         = {2008},
}

Altmetric
View in Altmetric
Web of Science
Times cited: