Quantification of urinary zwitterionic organic acids using weak-anion exchange chromatography with tandem MS detection

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• 作者：Michael Jason Bishop ; Brian S Crow ; Kasey D Kovalcik ; Joe George ; James A Bralley
• 关键词：Weak Anion Exchange ; Formiminoglutamic acid ; Pyroglutamic acid ; 5-hydroxyindoleacetic acid ; 2-methylhippuric acid ; LC/MS/MS ; Organic acids
• 刊名：Journal of Chromatography B, Analytical Technologies in the Biomedical and Life Sciences
• 出版年：2007
• 出版时间：1 April 2007
• 年：2007
• 卷：848
• 期：2
• 页码：303-310
• 全文大小：331 K

文摘

A rapid and accurate quantitative method was developed and validated for the analysis of four urinary organic acids with nitrogen containing functional groups, formiminoglutamic acid (FIGLU), pyroglutamic acid (PYRGLU), 5-hydroxyindoleacetic acid (5-HIAA), and 2-methylhippuric acid (2-METHIP) by liquid chromatography tandem mass spectrometry (LC/MS/MS). The chromatography was developed using a weak anion-exchange amino column that provided mixed-mode retention of the analytes. The elution gradient relied on changes in mobile phase pH over a concave gradient, without the use of counter-ions or concentrated salt buffers. A simple sample preparation was used, only requiring the dilution of urine prior to instrumental analysis. The method was validated based on linearity (r² ≥ 0.995), accuracy (85–115 % ), precision (C.V. < 12 % ), sample preparation stability (≤5 % , 72 h), and established patient ranges. The method was found to be both efficient and accurate for the analysis of urinary zwitterionic organic acids.