A comparison and column selection of Hydrophilic Interaction Liquid Chromatography and Reversed-Phase High-Performance Liquid Chromatography for detection of DNA methylation

Qualitative and Quantitative Analytical Techniques of Nucleic Acid Modification Based on Mass Spectrometry for Biomarker Discovery

“…In this one-step method, endonuclease from Serratia marcescens and Benzonase recognize single- and double-stranded DNA and RNA, and as the denaturing step is not essential, RNA samples could be hydrolyzed through the same workflow [ 43 ]. Directly hydrolyzing DNA or RNA into bases through heating under acidic conditions is also an alternative choice [ 45 , 46 , 47 ].…”

An on-line SPE-LC-MS/MS method for quantification of nucleobases and nucleosides present in biological fluids

“…HILIC-based methods were reported for the separation of nucleobases and nucleosides. [23][24][25][26] In this work, we have studied a new strategy for LC-MS quantication of nucleobases and nucleosides. It involves the conversion of these compounds to their respective ethenoderivatives with chloroacetaldehyde (CAA).…”

A relative quantitation method for measuring DNA methylation and hydroxymethylation using guanine as an internal standard

“…Nucleosides can be analyzed by chemiluminescence, capillary electrophoresis, thin layer chromatography, 15 and gas chromatography. 16 An alternative approach is high performance liquid chromatography, 13,17 which can be used for measuring the amount of nucleosides and free bases as well. Chromatographic separation is very difficult because of the high polarity of these compounds.…”

“…Chromatographic separation is very difficult because of the high polarity of these compounds. Mainly reversed phase (RP) chromatography 13,[17][18][19][20][21][22] and, in some cases, hydrophilic interaction liquid chromatography (HILIC) 17,23 techniques were established. Appropriate separation is challenging.…”

“…13,[23][24][25] This calibration model requires the measurement of the calibration sample points every time resulting in more sample preparation and a longer measurement time. Another popular approach is to determine the ratio of the molar quantities of the compound to the sum of cytosine-based molecules; 21,26,27 however, only a few articles have reported the use of deoxyguanosine (dG) for quantitation. 20,22 Our work on the identication of DNA methylation started several years ago.…”