For the dietary experiments where kyn was fed to the mice, kyn was synthesized via oxidation of oxindolylalanine as previously described (23 ). Product formation was monitored by mass spectrometry. Upon completion, the reaction was neutralized to pH 7 by the addition of HCl. Excess solvent was removed by rotary evaporation and kyn purified by solid phase extraction. The product was verified by NMR using a Varian multinuclear 400 MHz NMR spectrophotometer. The purity was assessed and the identity of the product confirmed by mass spectrometry (expected mass=208.2; actual mass=209.1) using an Agilent 1200 HPLC coupled to an Agilent 6120 quadrupole LC/MS. For the kyn injection studies, kyn was purchased commercially (Sigma).
Agilent 6120 quadrupole lc ms
Manufactured by Agilent Technologies
Sourced in United States
The Agilent 6120 quadrupole LC/MS is a liquid chromatography-mass spectrometry system. It provides accurate mass detection and quantification of analytes in complex samples.
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2 protocols using agilent 6120 quadrupole lc ms
1
Synthesis and Purification of Kynurenine
2
Purification and Characterization of Organic Compounds
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All the reactants, reagents, and solvents were obtained from commercial sources and used without further purification. Except where noted otherwise, the reactions were performed under an inert atmosphere of nitrogen gas using anhydrous solvents. The reactions were monitored using liquid chromatography–mass spectrometry (Agilent 1290 Infinity LC, Agilent 6120 quadrupole LC/MS, Agilent, Santa Clara, CA, USA). Flash chromatography was performed using the CombiFlash® system (Teledyne Isco, St. Lincoln, NE, USA). All nuclear magnetic resonance spectra were collected using a 600 MHz JNM-ECZ600R spectrometer (JEOL, Akishima, Tokyo). Chemical shifts (d in ppm) in the 1H spectra were reported relative to the residual solvent signals: 7.26 ppm for chloroform-d.
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