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A method for detecting small molecule metabolites in exosomes using mass spectrometry

A technology for detecting small molecule metabolites and mass spectrometry, which is applied in the field of detection of small molecule metabolites in exosomes by matrix-assisted laser desorption ionization mass spectrometry, achieving the effects of high salt tolerance, low preparation cost, and simple steps

Active Publication Date: 2020-01-21
浙江亿纳谱生命科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, traditional matrices are difficult to meet the needs of small molecule detection. A new type of matrix material that can be used for biological system detection and has certain anti-interference and salt tolerance needs to be developed urgently.

Method used

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  • A method for detecting small molecule metabolites in exosomes using mass spectrometry
  • A method for detecting small molecule metabolites in exosomes using mass spectrometry
  • A method for detecting small molecule metabolites in exosomes using mass spectrometry

Examples

Experimental program
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preparation example Construction

[0032] The preparation of the iron-containing oxide nanoparticle matrix comprises the following steps:

[0033] Step 1: dissolving ferric chloride and trisodium citrate in ethylene glycol solution;

[0034] Step 2: Add sodium acetate to the above mixed solution, and sonicate at room temperature for half an hour until the solution becomes a homogeneous system;

[0035] Step 3: The reaction will be carried out in a Teflon high-pressure reactor, and the reaction will be carried out at 150-195 degrees Celsius for more than 8 hours to form iron-containing oxide nanoparticles;

[0036] Step 4: repeatedly washing the iron-containing oxide nanoparticles obtained in step 3 with ethanol and deionized water, and finally drying at 62-70 degrees Celsius for use;

[0037] Step 5: Resuspend the iron-containing oxide nanoparticles in deionized water and use them as a matrix.

[0038] Characterization of the matrix:

[0039] The instruments used for characterization are: The instruments use...

Embodiment 1

[0043] Example 1: Detection of metabolic small molecules in exosomes extracted from serum

[0044] (1) Preparation of instruments and reagents: laser desorption ionization mass spectrometer, only mass spectrometry signals with a signal-to-noise ratio greater than 10 are used for analysis. Using AB SCIEX TOF / TOF TM 5800 mass spectrometer, Nd:YAG laser, wavelength 355nm. The pulse electric field delay extraction and reflection working mode is adopted, and the positive ion mode is used for detection. DataExplorer was used to observe, process, and analyze data, and only mass spectrometer signals with a signal-to-noise ratio greater than 10 were used for analysis.

[0045] (2) Preparation of iron-containing oxide nanoparticles.

[0046] (3) Take 1ml-10ml of serum, extract exosomes by ultra-high speed centrifugation at 15000-25000G for 4-10 hours, and prepare an exosome solution with a concentration of 0.1mg / ml-10mg / ml.

[0047] (4) The MALDI target plate was ultrasonically cle...

Embodiment 2

[0051] Example 2: Detection of metabolic small molecules in exosomes extracted from cells

[0052] (1) Preparation of instruments and reagents: laser desorption ionization mass spectrometer, only mass spectrometry signals with a signal-to-noise ratio greater than 10 are used for analysis. Using AB SCIEX TOF / TOF TM 5800 mass spectrometer, Nd:YAG laser, wavelength 355nm. The pulse electric field delay extraction and reflection working mode is adopted, and the positive ion mode is used for detection. DataExplorer was used to observe, process, and analyze data, and only mass spectrometer signals with a signal-to-noise ratio greater than 10 were used for analysis.

[0053] (2) Preparation of iron-containing oxide nanoparticles.

[0054] (3) Take 1ml-10ml cell solution (suitable for various types of cells), extract exosomes by ultra-high speed centrifugation at 15000-25000G for 4-10 hours, and prepare exosomes with a concentration of 0.1mg / ml-10mg / ml solution.

[0055] (4) The...

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Abstract

The invention discloses a method for detecting exosome small molecule metabolites by using mass spectrometry. The method comprises the following steps of preparing an instrument and a reagent; preparing an iron-containing oxide nanometer particle substrate; preparing an exosome sample; respectively taking the exosome samples diluted according to different proportions; performing sample application onto a target plate; performing drying at the room temperature; dispensing the iron-containing oxide nanometer particle substrate onto the exosome sample; performing drying at the room temperature; performing mass spectrometric detection; analyzing the mass spectrometric detection result to draw the conclusion. The method has the advantages that the micro-nano particle materials are used as the substrate; the qualitative and quantitative detection on molecules to be detected is fast and efficiently realized by using few biological samples in mass spectrometric analysis; meanwhile, the background interference and hotspot effects of the conventional substrate can be avoided; the detection on metabolism molecules in the exosome samples is realized.

Description

technical field [0001] The invention relates to the field of mass spectrometry detection, in particular to a method for detecting exosome small molecule metabolites by matrix-assisted laser desorption ionization mass spectrometry. Background technique [0002] Exosomes are small vesicles with a diameter of about 30-150nm and a density of 1.13-1.21g / m1. Exosomes are naturally present in bodily fluids, including blood, saliva, urine, and breast milk. Exosomes are membranous vesicles secreted by living cells derived from late endosomes (also known as multivesicular bodies). Exosomes have many important functions, such as performing protein transport functions, specifically targeting recipient cells, exchanging proteins and lipids, or triggering downstream signaling events. Exosomes also transport nucleic acids and participate in intercellular communication. In short, its protein, RNA and fat components are specific, and it carries some important metabolic small molecules, whi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/64
Inventor 钱昆刘彬
Owner 浙江亿纳谱生命科技有限公司
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