ABSTRACT
A highly sensitive (subpicomole level), structure-specific method of analysis has been developed for characterizing and quantitating the dinucleotide thymidylyl(3'----5')thymidine methyl phosphotriester by desorption chemical ionization tandem mass spectrometry. The methodology can be applied to related compounds such as the parent dinucleotide and employs either positive or negative ionization mass spectra or daughter spectra. The procedure allows specification of the site of methyl attachment.
Subject(s)
Dinucleoside Phosphates , Thymine Nucleotides/analysis , Chemical Phenomena , Chemistry , Mass SpectrometryABSTRACT
The coupling of a caesium ion source to a triple quadrupole mass spectrometer is described. The potential of this combination for examining thermally labile, non-volatile molecules, such as thiamine hydrochloride, is examined. Emphasis is placed on the advantages the various scanning modes of tandem mass spectrometry provide in ion structure elucidation and the investigation of desorption ionization mechanisms. Use of the caesium ion source for desorption of neutral molecules which are chemically ionized by an auxiliary gas is demonstrated. This procedure may be especially useful for examining non-volatile, non-ionic samples.
ABSTRACT
Quantitation of 7-methyldeoxyguanosine (m7dG) produced in the in vitro methyl methanesulfonate (MeMS) action on calf-thymus DNA is achieved by enzymatic degradation, liquid chromatographic separation and chemical ionization mass spectrometry. The total degree of methylation, measured by uptake of [14C]MeMS was 0.35%. Mass spectral analysis shows that m7dG constitutes 84% of the total methylated product. It is also shown that tandem mass spectrometry allows detection of m7dG, as the protonated base, down to 1 pmol level, suggesting that MS/MS analysis can be the method of choice in quantitation of the adducts of in vivo DNA modifications.
