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Application of silver nanosphere material in MALDI-TOF MS detection of small molecular metabolites

A metabolite, silver nanotechnology, applied in the field of mass spectrometry detection, can solve the problems of difficult analysis, matrix background interference, organic small molecule matrix rupture and association, etc., to overcome the matrix background interference phenomenon, improve detection sensitivity, and simplify detection difficulty. Effect

Inactive Publication Date: 2020-07-17
SICHUAN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, although there are many types of organic small molecule matrices used in MALDI-TOF MS, it is difficult to analyze small molecular weight (less than 10,000 Da) compounds. Severe matrix background interference phenomenon

Method used

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  • Application of silver nanosphere material in MALDI-TOF MS detection of small molecular metabolites
  • Application of silver nanosphere material in MALDI-TOF MS detection of small molecular metabolites
  • Application of silver nanosphere material in MALDI-TOF MS detection of small molecular metabolites

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1 prepares silver nanosphere

[0029] (1) Freshly prepared 48ml containing (2mM) NaBH 4 Mix with an aqueous solution of (4.28mM) trisodium citrate (TSC) and heat to 60°C, and stir in the dark for 30 minutes to ensure that the solution is uniform;

[0030] (2) Add 2 mL of AgNO dropwise to the resultant of step (1) 3 solution (1mM), then increase the temperature to 90°C, when the temperature reaches 90°C, use 0.1M NaOH solution to adjust the pH value of the solution to 10.5, and continue heating for 20 minutes until an obvious bright yellow color appears;

[0031] (3) The product obtained in step (2) was cooled at room temperature, then centrifuged at 12,000 r / min for 15 minutes to remove unreacted reducing agent, washed three times, then dispersed in deionized water, and finally stored at 4°C for future use.

[0032] The silver nanospheres prepared above are characterized and detected. The particle diameter of the silver nanospheres is 5nm, and its ultraviol...

Embodiment 2

[0034] Embodiment 2 prepares silver nanosphere

[0035] Take 250ml of AgNO3 (1.06mM), add 5ml of trisodium citrate (34mM) to it, then reflux and stir at 100°C, stir at a speed of 330 rpm for 45 minutes until the color of the reflux solution becomes muddy yellow, continue to stir , the solution was cooled to room temperature, filtered with a 0.2 μm membrane filter, washed with ultrapure water and purified by centrifugation, and finally resuspended with deionized water, and stored in a dark environment at 4°C for further use.

[0036] The silver nanospheres prepared above are characterized and detected, and the scanning electron microscope (SEM) figure of the silver nanospheres is shown in figure 2 , the UV-Vis absorption spectrum of silver nanospheres is shown in image 3 .

[0037] Depend on figure 2 It can be seen that the particle diameter of the silver nanosphere is 50nm, by image 3 It can be seen that the ultraviolet-visible light absorption peak of silver nanospher...

Embodiment 3

[0038] Example 3 Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry Detection

[0039] 1. Detection of small molecular substances in serum

[0040]Experimental group: drop 0.5uL of mixed amino acid sample (concentration is 200nM) on the polished steel target plate MTP384, and then add dropwise 1uL of the silver nanosphere material prepared in Example 1 after air-drying, the concentration of the silver nanosphere material is 1mg / ml, after being thoroughly dried, put it on the machine, and perform mass spectrometry detection on the Autoflex Max mass spectrometer (Bruker Daltonics Company of Bremen, Germany). 20 different points are randomly measured for each sample, and each point is randomly dotted 25 times by laser, so 500 satisfactory dots can be obtained.

[0041] Control group: the silver nanosphere material was replaced by α-cyano-4-hydroxyphenylacrylic acid (CHCA) matrix, and other processes were the same as the experimental group.

[0042] Th...

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Abstract

The invention provides an application of a silver nanosphere material in MALDI-TOF MS (Matrix Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry) detection of small molecular metabolites. The analysis method using the nano material as the MALDI-TOF MS matrix is suitable for mass spectrometry analysis of small molecules with molecular weight of less than 1000; the detection difficulty of a small molecule sample is greatly simplified; the detection sensitivity of MALDI-TOF MS of the small molecule sample is improved; and meanwhile, elimination of mass spectrum peak interference generated by an existing matrix and enhancement processing of a mass spectrum signal of the small molecule sample can be realized.

Description

technical field [0001] The invention belongs to the technical field of mass spectrometry detection, and in particular relates to the application of a silver nanosphere material in the detection of small molecular metabolites by MALDI-TOF MS. Background technique [0002] The application of metabolomics to clinical oncology research has become an area of ​​research in recent years as metabolic alterations can directly participate in transformation processes or support biological processes that enable tumor growth, and small molecule metabolites can be a unique source of cancer-specific information. hot spot. During this period, mass spectrometry detection methods are also playing an increasingly important role. [0003] Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) technology has become a useful technique in the analysis of biomolecules such as peptides, proteins and oligosaccharides. However, although there are many types of or...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/64B82Y40/00B82Y30/00
CPCB82Y30/00G01N27/64
Inventor 王云兵张华杨立张兴栋
Owner SICHUAN UNIV
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