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Liquid chromatography-mass spectrometry analysis method of phenylpropane metabolic pathway metabolite and application thereof

A liquid chromatography-mass spectrometry and analysis method technology, applied in the field of biochemical metabolism analysis, can solve the problems of many isomers, low qualitative reliability, weak detection sensitivity, etc.

Active Publication Date: 2021-09-17
QINGDAO AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the similar structure and many isomers of phenylpropane compounds, the commonly used detection methods have low qualitative reliability and weak detection sensitivity.

Method used

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  • Liquid chromatography-mass spectrometry analysis method of phenylpropane metabolic pathway metabolite and application thereof
  • Liquid chromatography-mass spectrometry analysis method of phenylpropane metabolic pathway metabolite and application thereof
  • Liquid chromatography-mass spectrometry analysis method of phenylpropane metabolic pathway metabolite and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1 Establishment of a liquid chromatography-mass spectrometry analysis method for phenylpropanoid metabolites:

[0028] 1. According to the synthesis sequence of the phenylpropanoid metabolic pathway, determine the metabolite category that needs to be analyzed and has a corresponding relationship: in order to determine the metabolite category that needs to be analyzed, the hydroxyl or methoxy group on the benzene ring is used as a replacement group during the synthesis process. The vertical classification standard, the horizontal classification standard is based on the compound types formed by the skeleton connection characteristics, and the structural periodic table of phenylpropanoid metabolites is established ( figure 2 ), a total of 103 metabolites were found for analysis.

[0029] 2. According to the structural characteristics of the metabolites, use the multiple reaction detection mode of the liquid mass analysis method to determine their analysis param...

Embodiment 2

[0031] Analysis of the content change trend of the phenylpropane metabolic pathway in the leaves of Ginkgo biloba in one year:

[0032] 1. Prepare samples for analysis

[0033] (1) Take 200 mg of Ginkgo biloba leaves to be tested in different months (stored at -80°C), and grind them fully under liquid nitrogen; (2) Add 1ml of 80% methanol and use ultrasonic-assisted extraction for 15 minutes at a temperature of 45°C; (3) Centrifuge at 12000g for 5 minutes, Take the supernatant extract; (4) add 1ml 80% methanol to the precipitate and repeat the above operation twice; (5) combine the three extracts, and use a nitrogen blower or a vacuum centrifuge to concentrate and volatilize the extract until there is no obvious Liquid; (6) add 80% methanol to dilute to 1ml, filter with 0.22μm organic membrane to remove impurities.

[0034] 2. Analyze the samples on the machine and obtain the analysis data

[0035] The samples were analyzed twice: 85 metabolites were analyzed in positive ion...

Embodiment 3

[0038] Example 3 Analysis of the change trend of phenylpropanoid metabolic pathways when soybean leaves are subjected to abiotic stress:

[0039] 1. Prepare samples for analysis

[0040] (1) Take 200 mg of soybean leaves to be tested (stored at -80°C) after salt treatment and drought treatment, and grind them fully under liquid nitrogen conditions; (2) Add 1ml of 80% methanol and use ultrasonic-assisted extraction for 15 minutes at a temperature of 45°C; (3 ) at 12000g for 5min, and take the supernatant extract; (4) add 1ml 80% methanol to the precipitate and repeat the above operation twice; Concentrate and evaporate until there is no obvious liquid; (6) add 80% methanol to make up to 1ml, and use a 0.22 μm organic filter membrane to remove impurities.

[0041] 2. Analyze the samples on the machine and obtain the analysis data

[0042] The samples were analyzed twice: 85 metabolites were analyzed in positive ion mode; 18 metabolites were analyzed in negative ion mode. (1) ...

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Abstract

The invention relates to the field of biochemical metabolism analysis, particularly relates that the content changes have correlation due to the fact that phenylpropane (allyl) acid metabolites, coumarin metabolites and flavonoids (collectively referred to as phenylpropane) metabolites have the front-back corresponding relation in the in-vivo synthesis process, the metabolic trend of phenylpropane compounds in an organic body is analyzed by quantitatively detecting the content changes of the metabolites. According to the invention, a qualitative identification and quantitative analysis method for 103 phenylpropane metabolites (including 62 isomers) is established by using a liquid chromatography-electrospray ionization-mass spectrometry technology, and is analyzed and applied to organisms such as gingko, soybean and the like. The invention aims to analyze the change trend of phenylpropane metabolic pathways by establishing a detection method which is high in sensitivity, accurate in qualitative analysis and simple to operate, and provides a new thought and a new method for researching a biological metabolic process.

Description

technical field [0001] The present invention relates to the field of biochemical metabolism analysis. Specifically, it utilizes the structural similarity between phenylpropanoids and flavonoid metabolites caused by the biosynthetic relationship, and quantitatively detects phenylacrylic (enzylic) acids and aromatics with the same substituent structure. Changes in the content of legumes and flavonoids metabolites determine the trend of changes in the metabolic pathways of phenylpropanoids, providing new ideas and methods for the study of biological metabolic processes. Background technique [0002] Phenylpropanoids (phenylpropanoids) are a class containing one or several C 6 -C 3 Compounds as the basic unit, including phenylpropene, phenylpropanol, phenylacrylic acid and its esters, coumarins, lignans, flavonoids, lignin, etc. These compounds play an important role in plant growth regulation and defense against disease attack and other life processes. Coumarin (coumarin) wa...

Claims

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Application Information

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IPC IPC(8): G01N30/06G01N30/88
CPCG01N30/06G01N30/88Y02A50/30
Inventor 孟杰周功克贺郭孔英珍王殿
Owner QINGDAO AGRI UNIV
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