Method for detecting tryptophan and metabolites and application

Abstract

The invention discloses a method for detecting tryptophan and metabolites and application, and relates to the technical field of metabolome analysis, and the method comprises the step of detecting the tryptophan and the metabolites which are extracted by a liquid-liquid extraction method and separated by a C18 chromatographic column by utilizing LC-MS / MS (Liquid Chromatography-Mass Spectrometry / Mass Spectrometry). A mobile phase adopted by liquid chromatography separation is composed of a water phase and an organic phase, the water phase is composed of formic acid with the concentration of 0.3%, 25mM ammonium formate and water, and the organic phase is composed of formic acid with the concentration of 0.3%, acetylacetone with the concentration of 0.25% and methanol. The reconstitution fluid is a methanol aqueous solution with the concentration of 40%. The mass spectrum scanning adopts an MRM technology; wherein the addition of acetylacetone solves the problem of peak tail removal, which cannot be solved by a conventional mobile phase. The existing method can only detect one of a serotonin metabolic pathway, a kynurenine metabolic pathway and an aromatic hydrocarbon metabolic pathway, but the method can detect 25 key metabolites of the three pathways at the same time, provides more clues for diagnosis, treatment and prevention of diseases, and is wider in application range.

Description

technical field [0001] The invention relates to the technical field of metabolome analysis, and more particularly to a method and application for detecting tryptophan and metabolites. Background technique [0002] In addition to its use in the synthesis of various proteins, tryptophan is also associated with various diseases such as neuronal function, autoimmune diseases, CNS diseases, infectious diseases, and cancer. There are three pathways for tryptophan metabolism, one is the serotonin pathway that forms tryptamine after decarboxylation by aromatic-L-amino acid decarboxylase (AADC); the other is the formation of serotonin (5-hydroxytryptamine) by tryptophan hydroxylase (TPH). -HT) aromatic hydrocarbon metabolism pathway; the third is converted to N-formylkynurenine (NFK) by three rate-limiting enzymes, IDO1, IDO2, and TDO, and NFK is under the action of kynurenine carboxamidase (AFMID). Kynurenine (Kyn) is formed, Kyn is converted to anthranilic acid (AA) by kynureninas...

AI Technical Summary

Problems solved by technology

[0004] A method and application for the detection of tryptophan and metabolites provided by the present invention, which adopts liquid-liquid extraction method to extract tryptophan and metabolites in the sample, detect after liquid phase separation, and use isotope internal standard The purpose of this method is to solve the problem that the existing technology cannot complete the detection of the entire pathway of tryptophan and metabolites, and there is a problem of peak tailing