Quantification of urinary zwitterionic organic acids using weak-anion exchange chromatography with tandem MS detection |
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Authors: | Bishop Michael Jason Crow Brian S Kovalcik Kasey D George Joe Bralley James A |
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Institution: | Metametrix Clinical Laboratory, 4855 Peachtree Ind. Blvd. Norcross, GA 30092, USA. mbishop@metametrix.com |
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Abstract: | 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>or=0.995), accuracy (85-115%), precision (C.V.<12%), sample preparation stability (
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