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Rapid determination of orotic acid in urine by a fast liquid chromatography/tandem mass spectrometric method
Author(s) -
Marca Giancarlo la,
Casetta Bruno,
Zammarchi Enrico
Publication year - 2003
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.980
Subject(s) - orotic acid , chemistry , chromatography , urine , isotope dilution , reproducibility , liquid chromatography–mass spectrometry , tandem mass spectrometry , high performance liquid chromatography , mass spectrometry , selected reaction monitoring , biochemistry
Quantification of orotic acid (uracil‐6‐carboxylic acid) in urine is an important tool to diagnose some inherited diseases, such as urea cycle disorder (OTCD) and hereditary orotic aciduria. New rapid analytical methods are necessary to provide high‐throughput orotic acid analyses. A new analytical method has been developed for the rapid analysis of orotic acid in urine by liquid chromatography coupled with ion spray tandem mass spectrometry (LC/MS/MS). After a sample dilution 1:20, the analysis was performed in the selected reaction monitoring mode in which orotic acid was detected through the transition m/z 155 to 111. The retention time was 3.9 min in a 4.5‐min analysis. Daily calibration between 0.5–5.0 μmol/L of orotic acid, corresponding to 10–100 μmol/L in urine before the 1:20 dilution, offered consistent linearity and reproducibility. Interassay coefficient of variance (c.v.) was 4.97% at a mean concentration of 10.99 μmol/L. The sensitivity and specificity of tandem mass spectrometry permitted a high volume of analyses of orotic acid. The sample preparation is simple, inexpensive and not time demanding. Copyright © 2003 John Wiley & Sons, Ltd.