title: Measurement of DNA methylation using stable isotope dilution and gas chromatography-mass spectrometry.
abstract: A simple, highly selective, and sensitive method has been developed to quantify methylation of DNA extracted from human peripheral blood mononuclear cells. Assay has been performed at nucleobases level. Cytosine and 5-methylcytosine DNA content has been detected by gas chromatography-mass spectrometry using [2-(13)C]cytosine and [2-(13)C]5-methylcytosine as internal standards. The methylation level has been calculated as 5-methylcytosine/total cytosine ratio. The working range selected on calibration curve, obtained by evaluation of standards and matrix-added standards measurements, is suitable for 5 microg DNA analysis. In this range, healthy human DNA methylation percentage is within 5-6%.
authors.affiliation: Dipartimento di Scienze e Tecnologie Alimentari e Microbiologiche (DiSTAM), International Center for the Assessment of Nutritional Status, Università degli Studi di Milano, 20133 Milan, Italy.
authors.forename: Antonella San
authors.initials: AS
authors.lastname: Romerio
authors.forename: Giovanni
authors.initials: G
authors.lastname: Fiorillo
authors.forename: Ileana
authors.initials: I
authors.lastname: Terruzzi
authors.forename: Pamela
authors.initials: P
authors.lastname: Senesi
authors.forename: Giulio
authors.initials: G
authors.lastname: Testolin
authors.forename: Alberto
authors.initials: A
authors.lastname: Battezzati
mesh_terms: D004247:DNA / Q000032:analysis* / Q000737:chemistry*; D019175:DNA Methylation; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D006801:Humans; D015394:Molecular Structure; D011865:Radioisotope Dilution Technique
chemical_list: D004247:DNA