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A high‐throughput monoamine oxidase inhibition assay using liquid chromatography with tandem mass spectrometry
Author(s) -
Yan Zhengyin,
Caldwell Gary W.,
Zhao Boyu,
Reitz Allen B.
Publication year - 2004
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.1415
Subject(s) - chemistry , chromatography , mass spectrometry , liquid chromatography–mass spectrometry , tandem mass spectrometry , monoamine oxidase , substrate (aquarium) , enzyme , organic chemistry , oceanography , geology
Abstract A highly efficient method utilizing liquid chromatography with tandem mass spectrometry (LC/MS/MS) was developed and employed for high‐throughput screening of compounds for monoamine oxidase (MAO) inhibition. The method used kynuramine as a common substrate for both MAO‐A and MAO‐B in incubations, and the 4‐hydroxyquinoline (4‐HQ) resulting from deamination of kynuramine followed by intramolecular condensation was analyzed using LC/MS/MS; formation of 4‐HQ was used as the marker of MAO activity to evaluate the effects of test compounds. Isocratic liquid chromatography was applied to reduce the run time to 2 min. Because of the high specificity and sensitivity of detection of 4‐HQ by LC/MS/MS, this method was able to use MAO enzymes at very low concentrations and to perform short incubations; as a result, consumable cost was minimized, and sample preparations were completely avoided. The inhibition data are highly reproducible, and the IC 50 values determined by the method are in good agreement with literature values. The results suggest that this method is very robust and can be used as a generic approach to screen for MAO inhibitors in drug discovery. Copyright © 2004 John Wiley & Sons, Ltd.