Premium
Analysis of organic acid markers relevant to inherited metabolic diseases by ultra‐performance liquid chromatography/tandem mass spectrometry as benzofurazan derivatives
Author(s) -
AlDirbashi Osama Y.,
Santa Tomofumi,
AlQahtani Khalid,
AlAmoudi Mohammed,
Rashed Mohamed S.
Publication year - 2007
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.3053
Subject(s) - chemistry , glutaric acid , chromatography , derivatization , mass spectrometry , reagent , selected reaction monitoring , liquid chromatography–mass spectrometry , tandem mass spectrometry , high performance liquid chromatography , urine , organic chemistry , biochemistry
Abstract We describe a new approach applicable to the determination of organic acids that serve as diagnostic markers for several inherited metabolic disorders. We utilized liquid chromatography/tandem mass spectrometry for analysis of organic acid derivatives of a recently described benzofurazan reagent. The derivatization step was necessary to obtain organic acid derivatives suitable for analysis by reversed‐phase liquid chromatography with high ionization efficiency for mass spectrometry in the positive‐ion mode. In this work, a group of related dicarboxylic acid markers containing five or six carbon atoms were analyzed and validation was performed for glutaric and 3‐hydroxyglutaric acids, the specific markers for glutaric acidemia type 1. Derivatization was achieved by reacting untreated urine with the derivatization reagent under mild conditions. The reaction mixture was analyzed on a C18 ultra‐performance liquid chromatography (UPLC) column (50 × 2.1 mm, 1.7 µm) and detected in the multiple reaction monitoring mode in 5 min. Calibration curves were linear up to at least 1000 µM with detection limits for glutaric and 3‐hydroxyglutaric acids of 0.025 and 0.02 µM, respectively (signal‐to‐noise ratio of 3). Intra‐day (n = 11) and inter‐day (n = 6) coefficients of variation were better than 11.2%. The assay was successfully applied to control (n = 134) and glutaric acidemia type 1 (n = 55) urine samples. Copyright © 2007 John Wiley & Sons, Ltd.