A hydrophilic interaction ultraperformance liquid chromatography (HILIC–UPLC) method for genomic DNA methylation assessment by UV detection
作者： Salvatore Sotgia , Angelo Zinellu , Elisabetta Pisanu , Luciano Murgia , Gerard Aime Pinna
DOI: 10.1007/S00216-010-3565-3
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摘要: A hydrophilic interaction chromatography-based method, in combination with 1.7 µm ethylene bridged hybrid particle packed column (100 mm × 2.1 I.D.) and ultraperformance liquid chromatography, has been developed to measure cytosine (C) methylcytosine (mC) order evaluate the extent of DNA methylation. Separation was achieved good resolution fairly short times (5.5 min) by using isocratic elution a mixture 97:3 (v/v) acetonitrile/10 mM ammonium acetate as mobile phase. The determination coefficients C mC were high (R 2 > 0.999) within range tested. %RSD for intraday interday respectively 2.2% 2.5% 3.5% 3.8% mC. limit detection 0.52 µM (0.52 fmol on-column) both while quantification 1.72 (1.72 smallest amount purified that yielded measurable level 10 µg. On whole, this method is simple, rapid, sensitive, precise.
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Joseph Sherma, Thin-Layer Chromatography-Densitometry Journal of AOAC International. ,vol. 88, pp. 1516- 1516 ,(2005) , 10.1093/JAOAC/88.5.1516
Joseph Deutsch, Aharon Razin, John Sedat, Analysis of 5-methylcytosine in DNA. I. Mass spectrometry. Analytical Biochemistry. ,vol. 72, pp. 586- 592 ,(1976) , 10.1016/0003-2697(76)90570-4
Jing Li, Chris Papadopoulos, Jimmy Xu, Growing Y-junction carbon nanotubes Nature. ,vol. 402, pp. 253- 254 ,(1999) , 10.1038/46214
Christina Dahl, Per Guldberg, DNA methylation analysis techniques. Biogerontology. ,vol. 4, pp. 233- 250 ,(2003) , 10.1023/A:1025103319328
Guo-Liang Xu, Timothy H. Bestor, Déborah Bourc'his, Chih-Lin Hsieh, Niels Tommerup, Merete Bugge, Maj Hulten, Xiaoyan Qu, James J. Russo, Evani Viegas-Péquignot, Chromosome instability and immunodeficiency syndrome caused by mutations in a DNA methyltransferase gene. Nature. ,vol. 402, pp. 187- 191 ,(1999) , 10.1038/46052
Susan J. Clark, Janet Harrison, Marianne Frommer, CpNpG methylation in mammalian cells Nature Genetics. ,vol. 10, pp. 20- 27 ,(1995) , 10.1038/NG0595-20
Antonella San Romerio, Giovanni Fiorillo, Ileana Terruzzi, Pamela Senesi, Giulio Testolin, Alberto Battezzati, Measurement of DNA methylation using stable isotope dilution and gas chromatography-mass spectrometry. Analytical Biochemistry. ,vol. 336, pp. 158- 163 ,(2005) , 10.1016/J.AB.2004.09.034
Kenneth C. Kuo, Roy A. McCune, Charles W. Gehrke, Rosemarie Midgett, Melanie Ehrlich, Quantitative reversed-phase high performance liquid chromatographic determination of major and modified deoxyribonucleosides in DNA Nucleic Acids Research. ,vol. 8, pp. 4763- 4776 ,(1980) , 10.1093/NAR/8.20.4763
J. Martin Bland, DouglasG. Altman, Statistical methods for assessing agreement between two methods of clinical measurement. The Lancet. ,vol. 327, pp. 307- 310 ,(1986) , 10.1016/S0140-6736(86)90837-8
Frank Lyko, Dirk Stach, Axel Brenner, Stephan Stilgenbauer, Hartmut Döhner, Michaela Wirtz, Manfred Wiessler, Oliver J. Schmitz, Quantitative analysis of DNA methylation in chronic lymphocytic leukemia patients ELECTROPHORESIS. ,vol. 25, pp. 1530- 1535 ,(2004) , 10.1002/ELPS.200305830