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Method for detecting indoleacetic acid and indolepropionic acid in serum and excrement

A detection method and technology of indole acetic acid, applied in the field of metabolite detection, can solve the problems of high price, complex matrix, and certain difficulty in removal, and achieve the effect of simple pretreatment process and low cost.

Pending Publication Date: 2021-11-09
SHANDONG ANALYSIS & TEST CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] At present, the analysis of indole acetic acid and indole propionic acid mainly includes liquid chromatography and liquid chromatography-mass spectrometry analysis. The sensitivity of liquid chromatography analysis is difficult to meet the requirements of trace analysis, so liquid chromatography-mass spectrometry is very popular.
Due to the complex matrix and many interfering substances of biological samples, such as serum and feces, so far, there is no liquid chromatography-mass spectrometry method for the qualitative and quantitative analysis of indole acetic acid and indole propionic acid in serum and feces.
[0005] In the prior art, derivatization treatment combined with liquid chromatography-mass spectrometry detection is generally adopted, and derivatization reagents are used to react with indole acetic acid and indole propionic acid to generate corresponding derivatives, thereby improving detection sensitivity and reducing detection difficulty , but most of the derivatization reagents are toxic and irritating substances, and the price is relatively expensive, and it is difficult to completely remove them later

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  • Method for detecting indoleacetic acid and indolepropionic acid in serum and excrement
  • Method for detecting indoleacetic acid and indolepropionic acid in serum and excrement
  • Method for detecting indoleacetic acid and indolepropionic acid in serum and excrement

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Embodiment 1

[0045] Determination of indole acetic acid and indole propionic acid in mouse serum

[0046] 1. Ultrasonic solvent extraction: Use a pipette gun to accurately pipette 100 μL of mouse serum into a 2 mL EP tube, add an appropriate volume of internal standards (indole acetic acid-d5 and indole propionic acid-d2), add 0.45 mL of extraction solution (methanol : water: formic acid = 15: 4: 1), sealed and ultrasonically extracted for 15 min, centrifuged at 10,000 rpm and 4°C for 10 min, and after taking the supernatant, add 0.45 mL of extract (methanol: water: formic acid = 15: 4 : 1) Repeat the extraction once, and combine the extracts.

[0047] 2. Enrichment and purification of solid-phase extraction cartridges: first activate the Oasis HLB (3cc / 60mg) solid-phase extraction cartridges with 4mL methanol and 4mL water, then accurately pipette 600μL extract solution for loading, add 4mL water for rinsing , and then eluted with 3 mL of eluent (0.1% formic acid in methanol), and collec...

Embodiment 2

[0054] Determination of indole acetic acid and indole propionic acid in mouse feces

[0055] 1. Homogeneous grinding + ultrasonic solvent extraction: Accurately weigh 0.05g mouse feces sample into a 2mL EP tube, add an appropriate volume of internal standards (indole acetic acid-d5 and indole propionic acid-d2), add 0.5mL extract (methanol: water: formic acid = 15:4:1), add grinding beads, grind at 4°C, ultrasonically extract for 15 minutes, centrifuge at 10,000 rpm for 10 minutes at 4°C, take the supernatant, and then add 0.5mL The extract (methanol: water: formic acid = 15:4:1) was repeatedly extracted once, and the extracts were combined.

[0056] 2. Enrichment and purification of solid-phase extraction cartridges: first activate the Oasis HLB (3cc / 60mg) solid-phase extraction cartridges with 4mL methanol and 4mL water, then accurately pipette 600μL extract solution for loading, add 4mL water for rinsing , and then eluted with 3 mL of eluent (0.1% formic acid in methanol),...

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Abstract

The invention relates to the field of metabolite detection, in particular to a method for detecting indoleacetic acid and indolepropionic acid in serum and excrement. Before liquid chromatography-mass spectrometry detection, a methanol-water-formic acid mixed solvent is adopted for extraction, and a solid-phase extraction small column is combined for purification and enrichment. The pretreatment process can significantly reduce matrix interference, and qualitative and quantitative detection of indoleacetic acid and indolepropionic acid in serum and excrement can be realized without derivatization treatment. The pretreatment method adopted by the invention is green and environment-friendly, and toxic and corrosive reagents are not introduced. The pretreatment process is simple, the cost is low, and the method has important significance for researching the action mechanism of indoleacetic acid and indolepropionic acid for protecting intestinal barriers and regulating host immunity.

Description

technical field [0001] The invention relates to the field of metabolite detection, in particular to a method for detecting indole acetic acid and indole propionic acid in serum and feces. Background technique [0002] The information disclosed in this background section is only intended to increase the understanding of the general background of the present invention, and is not necessarily taken as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to those skilled in the art. [0003] Indole derivatives such as indole acetic acid, indole propionic acid, indole formaldehyde, indole acetaldehyde and indole acrylic acid are small molecular compounds produced by the catabolism of tryptophan by intestinal microorganisms. source ligand. Aryl hydrocarbon receptor is an important ligand-activated transcription factor, which is widely expressed in various immune or non-immune cells, and participates in important biological proce...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06G01N30/08G01N30/14G01N30/72
CPCG01N30/02G01N30/06G01N30/08G01N30/14G01N30/72Y02A50/30
Inventor 宋月王泉博
Owner SHANDONG ANALYSIS & TEST CENT
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