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Matrix sample preparation method and matrix-assisted laser desorption ionization time-of-flight mass spectrometry method

A sample preparation and matrix technology, applied in the chemical field, can solve problems such as limited detection, and achieve the effects of wide application range, limited detection, and simple sample preparation methods

Active Publication Date: 2021-05-28
昆山聂尔精密仪器有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The object of the present invention is to provide a matrix sample preparation method and matrix-assisted laser desorption ionization time-of-flight mass spectrometry analysis method, to at least solve the problem of low molecular weight in the current use of organic acid matrix Problems with limited detection of analytes in MALDI-TOF mass spectrometry

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  • Matrix sample preparation method and matrix-assisted laser desorption ionization time-of-flight mass spectrometry method

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[0040] This embodiment provides a matrix sample preparation method, such as figure 1 Shown, described matrix sample preparation method comprises:

[0041] A1, HPLC grade acetonitrile and trifluoroacetic acid are mixed to obtain a standard solvent;

[0042] A2, adding 4-hydroxyl-3-nitrobenzonitrile in the standard solvent to obtain a matrix solution, wherein the structural formula of the 4-hydroxyl-3-nitrobenzonitrile is:

[0043]

[0044] A3, preparing the sample to be tested as a sample solution;

[0045] A4, after mixing the sample solution and the matrix solution at a volume ratio of 1:1, a mixed solution is obtained;

[0046] A5, spot the mixed solution on the target plate, and let it dry naturally to obtain the sample target plate.

[0047] It should be noted that the HPLC grade in the present invention means the purity of chemical reagents, meaning the purity of reagents that can be used in high performance liquid chromatography.

[0048] The matrix sample preparati...

Embodiment 1

[0064] This embodiment includes the substrate provided by the invention (4-hydroxyl-3-nitrobenzonitrile) and the existing conventional substrate (alpha-cyano-4-hydroxycinnamic acid (CHCA)) respectively to MRFA (a tetrapeptide ) for sample preparation and mass spectrometry results.

[0065] In this embodiment, the sample to be tested is MRFA, and the method for preparing the sample to be tested as a sample solution includes:

[0066] Dissolve MRFA in HPLC grade pure water to obtain an initial solution of MRFA with a concentration of 1000±20ppm;

[0067] HPLC grade pure water was added into the MRFA primary solution to obtain a MRFA sample solution with a concentration of 1±0.02 ppm.

[0068] specific:

[0069] 1. The preparation method of the matrix sample when the matrix provided in this embodiment is carried out to MRFA by mass spectrometry:

[0070] First, add 1000 μL of HPLC-grade acetonitrile and 1 μL of trifluoroacetic acid (TFA) into a 1.5 mL centrifuge tube, mix well...

Embodiment 2

[0090] This embodiment includes the matrix provided by the present invention (4-hydroxy-3-nitrobenzonitrile) and the existing conventional matrix (5-dimethoxy-4-hydroxycinnamic acid (SA)) to detect IgG protein respectively The sample preparation method and mass spectrometry results.

[0091] In this embodiment, the sample to be tested is an IgG protein, and the method for preparing the sample to be tested as a sample solution includes:

[0092] Dissolving trifluoroacetic acid in HPLC grade ultrapure water to obtain a trifluoroacetic acid solution with a concentration of 0.1±0.05%;

[0093] The IgG protein was dissolved in the trifluoroacetic acid solution to obtain an IgG protein sample solution, and the concentration of the IgG protein in the IgG protein sample solution was 1±0.02 mg / mL.

[0094] specific:

[0095] 1. The matrix sample provided by this embodiment is prepared by mass spectrometry detection of the IgG protein:

[0096] First, add 1000 μL of HPLC-grade aceton...

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Abstract

The invention provides a matrix sample preparation method and a matrix-assisted laser desorption ionization time-of-flight mass spectrometry method. The matrix sample preparation method comprises the steps: mixing HPLC-grade acetonitrile and trifluoroacetic acid, and obtaining a standard solvent; adding 4-hydroxy-3-nitrobenzonitrile into the standard solvent, and obtaining a matrix solution; preparing a to-be-detected sample into a sample solution; mixing the sample solution and the matrix solution according to a volume ratio of 1: 1 to obtain a mixed solution; and dripping the mixed solution on a target plate, and naturally performing drying to obtain the sample target plate. The matrix provided by the invention is wide in application range, is not only suitable for detection of small molecular compounds and polypeptide samples, but also suitable for detection of protein samples, in addition, the sample preparation method is simple and convenient during detection, and the problem that the detection is limited when an organic acid matrix is used for performing MALDI-TOF mass spectrometry on a low molecular weight analyte at present is solved.

Description

technical field [0001] The invention relates to the field of chemical technology, in particular to a matrix sample preparation method and a matrix-assisted laser desorption ionization time-of-flight mass spectrometry analysis method. Background technique [0002] Matrix-assisted laser desorption ionization mass spectrometry (Matrix-assisted laser desorption ionization mass spectrometry, MALDI-MS) is a mass spectrometry technology that came out in the late 1980s and developed rapidly. The ions produced by this ionization method are commonly detected by time offlight (TOF) detectors, so MALDI is often called matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) together with TOF. MALDI-TOF-MS technology has revolutionized the traditional mass spectrometry technology mainly used for the research of small molecular substances, and has since entered a new era of biological mass spectrometry technology development. The feature of this techniq...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/64G01N1/28
CPCG01N27/64G01N1/28
Inventor 韩永光
Owner 昆山聂尔精密仪器有限公司