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Ultra-performance liquid chromatography-tandem mass spectrometry quantitative profiling of tryptophan metabolites in human plasma and its application to clinical study
Ist Teil von
Journal of chromatography. B, Analytical technologies in the biomedical and life sciences, 2019-10, Vol.1128, p.121745-121745, Article 121745
Ort / Verlag
Netherlands: Elsevier B.V
Erscheinungsjahr
2019
Quelle
MEDLINE
Beschreibungen/Notizen
A sensitive, rapid and reliable ultra-performance liquid chromatography-tandem mass spectrometry (UPLC–MS/MS) method was developed and validated to assay tryptophan (TRP) and its nine metabolites, including kynurenine (KYN), kynurenic acid (KYNA), 3-hydroxykynurenine (3-HK), 3-hydroxyanthranilic acid (3-HAA), xanthurenic acid (XA), 5-hydroxytryptamine (5-HT), 5-hydroxyindoleacetic acid (5-HIAA), 3-indolepropionic acid (IPA) and 3-indoleacetic acid (IAA) in human plasma. Tryptophan-d5 (TRP-d5) and carbamazepine (CAR) were applied to the method quantification, where TRP-d5 was the corresponding internal standard (IS) for TRP and KYN, and CAR was the corresponding IS for the other analytes. Plasma samples were processed by deproteinisation with acetonitrile, followed by separation on an Acquity UPLC HSS T3 column by using gradient elution with 0.1% (v/v) formic acid in water and acetonitrile and detection by electrospray ionisation tandem mass spectrometry in positive ion multiple reaction monitoring (MRM) within a total run time of 5 min. The calibration ranges were 3–600 ng/mL for 3-HK, 1.5–300 ng/mL for 5-HT, 25–5000 ng/mL for KYN, 1–200 ng/mL for XA, 100–20,000 ng/mL for TRP, 5–1000 ng/mL for KYNA, 2–400 ng/mL for 3-HAA, 2.5–500 ng/mL for 5-HIAA and 10–2000 ng/mL for IAA and IPA. All intra- and inter-day analytical variations were acceptable. Matrix effect and recovery evaluation proved that matrix effect can be negligible, and sample preparation approach was effective. The newly developed method can simultaneously determine a panel of TRP metabolites and was successfully applied in the clinical study characterising TRP metabolism in healthy volunteers.
•Quantitative profiling of 10 tryptophan metabolites involved in kynurenine, serotonin and indole-3-pyruvate/tryptamine pathways by UPLC-MS/MS in human plasma.•The method is sensitive, precise and time-efficient, enabling a high-throughput assay.•Successfully evaluating the association between the biochemical indicators and tryptophan metabolites’ concentrations in 100 healthy individuals.