Gas chromatographic-mass spectrometric determination of 5-hydroxymethyluracil in human urine by stable isotope dilution.
A method for the determination of 5-hydroxymethyluracil in urine is described. 5-Hydroxymethyluracil was extracted by reversed-phase chromatography and quantified by gas chromatography-mass spectrometry as tert.-butyldimethylsilyl derivative. Since natural 5-hydroxymethyluracil contained ca. 22% of M + 2 species, an internal standard consisting of [1,3-15N2,5-2H2]hydroxymethyluracil was used to correct losses during extraction, evaporation and derivatization. Between-run precision of this method was 7.79%, and concentrations as low as 1.87 nM could be measured. This sensitivity and precision could not be obtained with trimethylsilyl derivatives.
Laboratoire de Biochimie C, C.H.U., Grenoble, France.
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Incardona
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Boujet
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Cadet
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Ducros
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D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D007201:Indicator Dilution Techniques; D017646:Organosilicon Compounds; D010432:Pentoxyl / Q000031:analogs & derivatives* / Q000652:urine; D012825:Silicon / Q000032:analysis
D017646:Organosilicon Compounds; C009660:t-butyldimethylsilyl compounds; C008055:5-hydroxymethyluracil; D010432:Pentoxyl; D012825:Silicon