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2 protocols using d5 kynurenic acid

1

Comprehensive Tryptophan Metabolite Analysis

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Isotopically labeled internal standards d5-tryptophan (TRPd, 98.8%), d4-serotonin (5HTd, 98.7%), and d5-kynurenic acid (KINAd, 99.2%) were purchased from CDN isotopes (Quebec, Canada) while 13C6-nicotinamide (NAmC, 99.4%) was purchased from Sigma Aldrich (Milan, Italy). Analytical standards for tryptophan (TRP), kynurenine (KYN), 5-hydroxy-tryptophan (5-HTP), serotonin (5-HT), tryptamine (Tryp), kynurenic acid (KYNA), quinaldic acid (QA), xanthurenic acid (XA), 3-hydroxyanthranilic acid (3-HAA), 5-hydroxyindoleacetic acid (5-HIAA), nicotinic acid (NA), quinolinic acid (QuiA), and nicotinamide (NAm) have been purchased from Sigma Aldrich (Milan, Italy). LC-MS-grade solvents (acetonitrile, methanol, chloroform, isopropanol), and suprapure trifluoroacetic acid (TFA) were purchased from Romil. TRIzol™ reagent was obtained from Thermo Fisher Scientific.
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2

Quantification of Neurotransmitters and Metabolites

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Anthranilic acid, dopamine hydrochloride, epinephrine hydrochloride, L-glutamic acid, GABA, kynurenic acid, Lkynurenine, metanephrine, serotonin hydrochloride and Ltryptophan were purchased as powders from Sigma Aldrich (Sydney, Australia). D3-Dopamine, D5-glutamic acid, D6-GABA, D5-kynurenic acid and D3-metanephrine were supplied by CDN Isotopes (Quebec, Canada). HPLC grade acetonitrile was purchased from Honeywell (Sydney, Australia). Formic acid and acetic acid were supplied by RCI Labscan (Bangkok, Thailand). Three millilitre syringes were purchased from Beckton Dickinson (Sydney, Australia) and 0.22 μm hydrophilic syringe filters were supplied by Micro-Analytix Pty Ltd. (Taren Point, NSW, Australia).
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