Separation and identification of the 4-hydroxyantipyrine sulphoconjugate.
In a previous study we observed, during separation of total antipyrine metabolites by high-performance liquid chromatography and after enzymatic hydrolysis, an unidentified peak corresponding to an ionic compound with pyrazolinone features. In the present study, this compound was identified as the 4-hydroxyantipyrine sulphoconjugate, and its structure was definitively confirmed by gas chromatographic-mass spectrometric analysis and by the use of pure synthetic substance. We also demonstrated the inhibitory effect of sodium metabisulphite, a necessary preservative of urinary samples, on hydrolysis of this conjugate in the presence of sulphatases from Helix pomatia or Aerobacter aerogenes. This inhibitory effect makes it impossible to perform a global assay of antipyrine metabolites after enzymatic or chemical hydrolysis and confirms the value of direct assay of the 4-hydroxyantipyrine sulphoconjugate.
Moreau
J
J
Département de Biochimie, Centre Hospitalier de Versailles, Le Chesnay, France.
Palette
C
C
Cordonnier
P
P
Naline
E
E
Advenier
C
C
Pays
M
M
D000328:Adult; D000983:Antipyrine / Q000031:analogs & derivatives* / Q000378:metabolism / Q000652:urine; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002849:Chromatography, Gas; D002851:Chromatography, High Pressure Liquid; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D006868:Hydrolysis; D007202:Indicators and Reagents; D013058:Mass Spectrometry; D013429:Sulfatases / Q000378:metabolism; D013447:Sulfites
D007202:Indicators and Reagents; D013447:Sulfites; C007983:4-hydroxyantipyrine; C005200:sodium metabisulfite; D013429:Sulfatases; D000983:Antipyrine