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A method for detection of sphingolipids in zebrafish brain tissue based on uhplc-qtrap MS

A technique for brain tissue, zebrafish, applied in analytical chemistry

Active Publication Date: 2021-07-06
INST OF QUALITY STANDARD & TESTING TECH FOR AGRO PROD OF CAAS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] At present, the quantitative detection method of sphingolipids in zebrafish brain tissue has not been reported, so it is of great practical significance to develop a rapid and sensitive detection method for sphingolipids in zebrafish brain tissue

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  • A method for detection of sphingolipids in zebrafish brain tissue based on uhplc-qtrap MS
  • A method for detection of sphingolipids in zebrafish brain tissue based on uhplc-qtrap MS
  • A method for detection of sphingolipids in zebrafish brain tissue based on uhplc-qtrap MS

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preparation example Construction

[0040] In some embodiments, the preparation method of the zebrafish brain tissue protein comprises:

[0041] 1) lyse brain tissue and precipitate protein by protein precipitant;

[0042] 2) adding chloroform to vibrate and stratify, and take the chloroform layer;

[0043] Optional, repeat step 2);

[0044] 3) Blow dry the chloroform layer with nitrogen.

[0045] In some embodiments, the protein precipitation agent is selected from methanol and / or acetonitrile.

[0046] In some embodiments, the preparation method of the zebrafish brain tissue protein also includes:

[0047] Reconstitute with methanol, and pass through a 0.20-0.24 μm microporous membrane, or 0.22 μm.

[0048] Since there are many impurities in animal tissues and the physical and chemical properties of organic solvents are quite different, sample pretreatment is a very critical part. The workload and error sources of pre-processing accounted for more than 60% of the entire detection process. The focus is on ...

Embodiment 1

[0058] Detection of sphingolipids in zebrafish brain tissue

[0059] 1. Preparation of physical samples: Take 5 fish brains (about 20 mg) and perform a blank subtraction and addition recovery test. At the same time, set a blank control and add samples, and add concentrations of 5 μg / kg, 10 μg / kg and 50 μg / kg. Evenly, fully stand. Add 180 μL of methanol to the 2 mL centrifuge tube containing the zebrafish brain tissue sample, and then grind it thoroughly on ice. Transfer the uniformly ground brain tissue to a 5mL centrifuge tube, then add 180 μL of methanol to wash the tube wall of the 2mL centrifuge tube, and transfer it to a 5mL centrifuge tube as well. Add 1.2 mL of chloroform, vortex for 1 min, and ultrasonically extract for 3 min. After the sonication, 240 μL of water was added and shaken to make it appear stratified. The chloroform layer was taken and extracted after repeated addition of chloroform. Then the chloroform layers obtained twice were combined, and blown dr...

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Abstract

The present invention relates to the field of analytical chemistry and the field of lipidomics research, in particular, to a method for detecting sphingolipids in zebrafish brain tissue based on UHPLC-QTrap MS, wherein the UHPLC is used to determine the extraction of zebrafish brain tissue protein When liquid, carry out gradient elution with mobile phase A and mobile phase B; Described mobile phase A is the formic acid aqueous solution of 0.3v / v%~0.5v / v%; Described mobile phase B is that volume ratio is 40:(50 ~70) 0.07v / v%~0.13v / v% formic acid in water: isopropanol. The method can realize simultaneous quantitative and qualitative detection of 46 sphingolipid substances in zebrafish brain tissue, and is time-consuming and low-cost.

Description

technical field [0001] The invention relates to the fields of analytical chemistry and lipidomics research, in particular to a method for detecting sphingolipids in zebrafish brain tissue based on UHPLC-QTrap MS. Background technique [0002] Zebrafish (English name: Danio rerio) is a commonly used model organism for toxicological evaluation. Zebrafish have natural, intrinsic advantages over other model organisms. First of all, zebrafish are easy to obtain and raise, and can be cultured on a large scale in the laboratory. Secondly, the zebrafish passage cycle is short, which can meet the experimental requirements. Most importantly, zebrafish genes are 87% similar to human genes. Based on the above advantages, zebrafish is often used as a model organism for toxicological evaluation of pesticides, veterinary drugs, persistent organic pollutants and new environmental pollutants. [0003] Sphingolipids (sphingolipids) are a class of amphoteric lipids containing a sphingosine...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02G01N30/72
CPCG01N30/02G01N30/72
Inventor 邱静宋月钱永忠
Owner INST OF QUALITY STANDARD & TESTING TECH FOR AGRO PROD OF CAAS
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