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Application of High Performance Liquid Chromatography/Mass Spectrometry with Electrospray Ionization to the Detection of DNA Nucleosides in Ancient Seeds
Author(s) -
O'Donoghue Kerry,
Brown Terence A.,
Carter James F.,
Evershed Richard P.
Publication year - 1996
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/(sici)1097-0231(19960331)10:5<495::aid-rcm492>3.0.co;2-d
Subject(s) - chemistry , chromatography , electrospray ionization , ammonium acetate , mass spectrometry , formic acid , electrospray , direct electron ionization liquid chromatography–mass spectrometry interface , deoxyribonucleosides , high performance liquid chromatography , dna , chemical ionization , ion , ionization , organic chemistry , biochemistry
Liquid chromatography/mass spectrometry (LC/MS) with electrospray ionization has been used to analyse the nucleoside components of DNA extracted from preserved plant remains. Optimum results were obtained by use of an isocratic ammonium acetate/acetonitrile mobile phase with post‐column addition of 50% v/v aqueous formic acid. Authentic DNA nucleosides gave simple spectra displaying protonated molecular ions, [M+H] + , and base ions, [BH 2 ] + , carrying the bulk of the ion current. Selected ion monitoring (SIM) of these characteristic ions was employed to screen for the presence of deoxyribonucleosides in enzymatic hydrolysates of DNA isolated from ancient seeds. This use of mass chromatography with co‐injection analyses confirmed the presence of 2′‐deoxycytidine, 2′‐deoxyadenosine and 2′‐deoxyguanosine in 1400 year‐old desiccated radish ( Raphanus sativus ) and barley ( Hordeum sp. ) seeds. Detection of thymidine could not be confirmed due to a low response of this nucleoside under the ionization conditions used. The use of LC/MS‐SIM overcomes problems of low volatility and co‐eluting contaminants previously seen with GC/MS‐SIM of DNA from ancient seeds.