Electrophore-labeling and alkylation of standards of nucleic acid pyrimidine bases for analysis by gas chromatography with electron-capture detection.
The pyrimidine bases cytosine, uracil and thymine, along with some analogues, are electrophore-labeled either with pentafluorobenzoyl chloride (PFBC), pentafluorophenylsulfonyl chloride (PPSC), or heptafluorobutyric anhydride. Subsequent alkylation is most successful for PFB-cytosine, PPS-uracil, and PPS-thymine. These same alkylated compounds also have the highest aqueous stability and respond most strongly by gas chromatography-electron-capture detection. One of these derivatives, determined to be N4-PFB-1,3-dimethylcytosine by authentic synthesis, and its 5-methyl analogue, can be detected with good precision down to the 100-fg level. Poor reproducibility is encountered at the 10-fg level.
D000478:Alkylation; D055598:Chemical Phenomena; D002621:Chemistry; D002849:Chromatography, Gas / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002853:Chromatography, Liquid / Q000379:methods; D002855:Chromatography, Thin Layer / Q000379:methods; D011743:Pyrimidines / Q000032:analysis*; D012015:Reference Standards
D011743:Pyrimidines