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Determination of N-7-[ 2H 3]methyl guanine in rat urine by gas chromatography-mass spectrometry following administration of trideuteromethylating agents or precursors

Authors
  • Shuker, David E.G.
  • Bailey, Eric
  • Gorf, Susan M.
  • Lamb, John
  • Farmer, Peter B.
Type
Published Article
Journal
Analytical Biochemistry
Publisher
Elsevier BV
Publication Date
Jan 01, 1984
Volume
140
Issue
1
Pages
270–275
Identifiers
DOI: 10.1016/0003-2697(84)90164-7
Source
Elsevier
Keywords
License
Unknown

Abstract

A gas chromatographic-mass spectrometric method has been developed for the determination of N-7-[ 2H 3]methyl guanine in urine in the presence of large natural levels of N-7-methyl guanine. Urine is fractionated on heptanesulfonic acid-treated C-18 Sep-pak cartridges, followed by derivatization to give a volatile N-heptafluorobutyryl- O 6-2,3,4,5,6-pentafluorobenzyl derivative which is separated on an SE52 fused silica capillary column. Using N-7-ethyl guanine as an internal standard, the total amount of N-7-methyl guanine is determined by gas chromatography-flame ionization detection. The percentage of N-7-[ 2H 3]methyl guanine is then measured by gas chromatography-mass spectrometry, enabling the amount of deuterated base to be determined. Preliminary experiments with [ 2H 3]methyl methanesulfonate in rats showed measurable excretion of N-7-[ 2H 3]methyl guanine. 4-(Di[ 2H 3]methylamino)antipyrine alone gave no detectable amount of alkylated base, but coadministration of nitrite resulted in excretion of deuterated N-7-methyl guanine.

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