Ion stability of nucleic acids in infrared matrix-assisted laser desorption/ionization mass spectrometry.
Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) with infrared laser light of a wavelength of 2.94 microns has been used for the analysis of nucleic acids. Spectra of oligodeoxynucleotides up to 26 nucleotides, oligothymidylic acids up to 100 nucleotides as well as different synthetic RNA oligomers and RNA transcripts up to 104 nucleotides are presented. A main problem in the analysis of oligodeoxynucleotides was found to be related to the loss of bases. The stability of oligothymidylic acids as opposed to oligodeoxynucleotides containing all four bases indicates that the loss of bases is correlated with A, C and G protonation which decreases the stability of the N-glycosidic bond. Experiments indicate that the breakage of the N-glycosidic bond probably occurs during the desorption process due to proton transfer from the phosphodiester groups to the ionizable bases. RNA displayed a significantly higher stability in MALDI-MS due to the presence of a 2'-OH group. Consequently, signals of RNA transcripts with a length of up to 142 nucleotides could be detected by MALDI-MS. Technical details of the method, including the distribution of positive counterions on the phosphodiester backbone, the upper mass limit and mass accuracy are discussed along with a number of potential analytical applications.
Nordhoff
E
E
Institute for Medical Physics and Biophysics, University of Münster, Germany.
Cramer
R
R
Karas
M
M
Hillenkamp
F
F
Kirpekar
F
F
Kristiansen
K
K
Roepstorff
P
P
D001483:Base Sequence; D004247:DNA / Q000032:analysis / Q000737:chemistry; D004355:Drug Stability; D006863:Hydrogen-Ion Concentration; D007834:Lasers; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D009696:Nucleic Acids / Q000032:analysis* / Q000737:chemistry; D012313:RNA / Q000032:analysis / Q000737:chemistry
D009696:Nucleic Acids; D012313:RNA; D004247:DNA