A sensitive high-performance liquid chromatographic procedure with fluorometric detection for the analysis of decarboxylated S-adenosylmethionine and analogs in urine samples.
A highly sensitive HPLC method for the determination of decarboxylated S-adenosylmethionine (dc-SAM) by fluorometric detection was developed. The reaction of dc-SAM and its analogs with chloroacetaldehyde leads to the corresponding 1,N6-etheno derivatives. These highly fluorescent derivatives were fully characterized through their proton nuclear magnetic resonance spectra and/or mass spectra. This derivatization procedure has been applied to the analysis of dc-SAM in rat and human urine. After a simple cation exchange column prepurification, the urine extracts were derivatized with chloroacetaldehyde and analyzed by reversed-phase HPLC with fluorometric detection. The method allowed the determination of subpicomole amounts of dc-SAM and was shown to be highly reproducible with the use of decarboxylated S-adenosylethionine as internal standard. The application of the method to the analysis of urine of rats treated with MDL 72175, a potent ornithine decarboxylase inhibitor, showed that the dc-SAM levels increased in a dose-related fashion.
D000480:Alkynes; D000818:Animals; D055598:Chemical Phenomena; D002621:Chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003653:Decarboxylation; D003959:Diamines / Q000494:pharmacology; D006801:Humans; D008297:Male; D065108:Ornithine Decarboxylase Inhibitors; D051381:Rats; D012436:S-Adenosylmethionine / Q000031:analogs & derivatives* / Q000652:urine; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet
D000480:Alkynes; D003959:Diamines; D065108:Ornithine Decarboxylase Inhibitors; C012702:S-adenosyl-3-methylthiopropylamine; D012436:S-Adenosylmethionine; C039613:6-heptyne-2,5-diamine