Global DNA methylation measured by liquid chromatography-tandem mass spectrometry: analytical technique, reference values and determinants in healthy subjects

Abstract

This LC-ESI-MS/MS method is easy to perform and offers reproducibility, selectivity and sensitivity for studying DNA methylation. The method allows a sample throughput of approximately 200 samples/week. The MTHFR C677T genotype influences age-related changes in DNA methylation.

DNA methylation in white blood cells

“…Serum folate, red blood cell 5-methyltetrahydrofolate, red blood cell non-methylfolate and red blood cell total folate were not associated with 5-mC content in blood. 81 Another study did not observe any association between 5-mC content in blood and nutrients including folate, vitamin B1, B2, B3, B12 and total protein. 18 Individuals with lower dietary folate showed no difference in LINE-1 methylation compared with individuals with higher dietary folate intake.…”

Identification of DNMT1 Selective Antagonists Using a Novel Scintillation Proximity Assay

“…5-AZA (6 M) was used as a positive control. Global methylation levels were determined, following DNA hydrolysis, by an LC-MS/MS method as described previously (26).…”

“…An LC-MS/MS-based genomic methylation assay was employed (26). This assay relies on acid hydrolysis of DNA to yield free cytosine and 5-methylcytosine followed by separation on a C18 reversed phase column using the ion-pairing agent nonafluoropentanoic acid and quantification using stable isotope dilution.…”

Alcohol abuse and cigarette smoking are associated with global DNA hypermethylation: Results from the German Investigation on Neurobiology in Alcoholism (GINA)

“…Global DNA methylation was measured by ultra-high performance liquid chromatography (UHPLC) tandem mass spectrometry with a method adapted from the work done by Kok and coworkers (Kok et al, 2007). In short, 2 µg of DNA were hydrolyzed using formic acid.…”