Kit and method for detecting tryptophan and metabolites thereof based on UPLC-MS/MS

A technology of tryptophan and metabolites, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of few types of metabolites, cumbersome pretreatment, high blood consumption, etc., and achieve simple pretreatment operation and low plasma consumption Low, the effect of improving accuracy

Pending Publication Date: 2021-09-17
广州市红十字会医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The purpose of the present invention is to provide a method for detecting human plasma tryptophan and its Accurate, convenient and high-throughput kits and methods for its metabolites

Method used

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  • Kit and method for detecting tryptophan and metabolites thereof based on UPLC-MS/MS
  • Kit and method for detecting tryptophan and metabolites thereof based on UPLC-MS/MS
  • Kit and method for detecting tryptophan and metabolites thereof based on UPLC-MS/MS

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Embodiment

[0078]The following examples are used herein to demonstrate preferred embodiments of the present invention. Those skilled in the art will appreciate that the techniques disclosed in the following examples represent techniques discovered by the inventors that can be used to implement the present invention, and thus can be regarded as preferred solutions for implementing the present invention. However, those skilled in the art should understand from this specification that many modifications can be made to the specific embodiments disclosed herein and still obtain the same or similar results, without departing from the spirit or scope of the present invention.

[0079] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs, the public references herein and the materials to which they refer are incorporated by reference .

[0080] Those skilled in th...

Embodiment 2

[0126] Example 2 Selective verification of the method established in Example 1

[0127] The selectivity of the method established in Example 1 was tested for the selectivity of the method established in Example 1 by analyzing the responses at the peaks of the analyte and internal standard with the lowest limit of quantification in blank plasma from 6 different individual sources.

[0128] The results show that the peak area of ​​the blank plasma analyte does not exceed 20.0% of the lower limit of quantification and the internal standard does not exceed 5.0%, indicating that there are no interfering components, and the method of the present invention has good selectivity.

Embodiment 3

[0129] Example 3 Linearity verification of the method established in Example 1

[0130] Taking the mass concentrations of the 8 analytes as the abscissa x, and the ratio of the peak area of ​​the analyte to the internal standard as the ordinate y, the linear regression analysis was performed by the weighted least squares method (the weight coefficient was 1 / x). 2 ), select the mixed internal standard solution containing tryptophan-d5 and carbamazepine. According to the chromatographic behavior and ionization efficiency of the two internal standards and analytes in the method of the present invention, tryptophan-d5 was used as the internal standard for kynurenine and tryptophan, carbamazepine was used as the internal standard for other analytes, and Plasma concentrations of tryptophan and its metabolites were calculated by double internal standard method.

[0131] The results show that the 10 analytes have good linearity in the following range, and the correlation coefficient ...

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Abstract

The invention discloses a kit and a method for simultaneously detecting tryptophan and metabolites thereof in a biological sample based on ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), which belong to the technical field of UPLC-MS/MS detection. The kit comprises standard substances of tryptophan and metabolites of the tryptophan, and the tryptophan and the metabolites of the tryptophan comprise 3-hydroxyl kynurenine, 5-hydroxytryptamine, kynurenine, yellow uric acid, tryptophan, kynurenic acid, 3-hydroxyl o-aminobenzoic acid, 5-hydroxyl indoleacetic acid, 3-indoleacetic acid and 3-indolepropionic acid. According to the method, the tryptophan and the metabolites of the tryptophan are detected by utilizing UPLC-MS/MS, and a standard curve is established according to a peak area, so that the contents of the tryptophan and the metabolites of the tryptophan are obtained. The method for detecting the tryptophan and the metabolites of the tryptophan is simple to operate, has the characteristics of high sensitivity, good selectivity and the like, can accurately and quickly detect the tryptophan and the metabolites of the tryptophan at the same time, and has wide application value.

Description

technical field [0001] The invention relates to the technical field of ultra-high performance liquid chromatography tandem mass spectrometry (UPLC-MS / MS) detection, in particular to a kit and method for detecting tryptophan and its metabolites based on UPLC-MS / MS. Background technique [0002] Tryptophan (TRP) is one of the essential amino acids in the human body, which can play an important role in protein biosynthesis and metabolic regulation network in vivo as the precursor of many important biologically active molecules. According to research, diseases such as cancer, coronary heart disease, Alzheimer's disease, and irritable bowel syndrome are all related to the metabolic disorder of tryptophan. Intake of tryptophan is mainly metabolized in the body through the following three pathways: kynurenine (KYN), serotonin (Serotonin) pathway and bacterial degradation pathway. [0003] About 95% of tryptophan is metabolized through the kynurenine pathway and converted into a se...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/34G01N30/86
CPCG01N30/02G01N30/06G01N30/34G01N30/8634G01N2030/045
Inventor 张述耀陈云黄逸辉江红陈旺傅永锦廖巍黄小珊
Owner 广州市红十字会医院
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