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Simultaneous Quantification of Selected Kynurenines Analyzed by Liquid Chromatography-Mass Spectrometry in Medium Collected from Cancer Cell Cultures
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Chapters
- 00:04Introduction
- 00:573-Nitrotyrosine (3NT), Kynurenine (Kyn), 3-Hydroxykynurenine (3HKyn), 3-Hydroxyanthranilic Acid (3HAA), Xanthurenic Acid (XA) and Charcoal Treated Culture Medium Preparation
- 01:58Calibration Solution Preparation
- 03:42Liquid Chromatography-Mass Spectrometry (LC-MS) System Setu
- 04:20Calibration Curve Construction
- 04:55Sample Collection
- 05:33Sample Preparation for LC-MS Analysis
- 06:30Protein Content Assessment and Data Normalization
- 08:12Results: Representative LC-MS Data of Selected KYNURENINES Determination
- 09:57Conclusion
Described here is a protocol for the determination of four different tryptophan metabolites generated in the kynurenine pathway (kynurenine, 3-hydroxykynurenine, xanthurenic acid, 3-hydroxyanthranilic acid) in the medium collected from cancer cell cultures analyzed by liquid chromatography coupled with a single quadrupole mass spectrometry.
