CN110146586B - MALDI-MS method for detecting small molecule metabolite by 1,1 '-binaphthyl-2, 2' -diamine and application - Google Patents
MALDI-MS method for detecting small molecule metabolite by 1,1 '-binaphthyl-2, 2' -diamine and application Download PDFInfo
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Abstract
MALDI-MS method for detecting small molecule metabolite by 1,1 '-binaphthyl-2, 2' -diamine and application thereof. The method comprises the following steps: 1) transferring the frozen section made of the fresh biological tissue onto a conductive glass slide, and performing vacuum pumping treatment to obtain a tissue section; 2) preparing a solution from a matrix and a solvent by taking 1,1 '-binaphthyl-2, 2' -diamine as the matrix, spraying the solution onto the tissue slice obtained in the step (1), and performing MALDI mass spectrometry imaging analysis; and then carrying out targeted MALDI mass spectrum data extraction on different kinds of small molecule metabolites in the biological tissue to obtain the ion intensity value of the small molecule metabolites. The invention takes 1,1 '-binaphthyl-2, 2' -diamine as a matrix to carry out MALDI-MS analysis on biological tissues, not only has low background interference, but also obviously improves the detection sensitivity of small molecule metabolites, and simultaneously can realize high coverage analysis of various metabolites in the biological tissues.
Description
Technical Field
The invention belongs to the technical field of mass spectrometry detection, and particularly relates to application of 1,1 '-binaphthyl-2, 2' -diamine as a MALDI-MS matrix.
Background
The information disclosed in this background of the invention is only for enhancement of understanding of the general background of the invention and is not necessarily to be taken as an acknowledgement or any form of suggestion that this information forms the prior art already known to a person skilled in the art.
The small molecule metabolite (m/z <1000) is a product of the combined action of the organism metabolism and the external environment, and compared with genes and proteins, the small molecule metabolite can more directly and accurately reflect the state of a biological system. Researches show that the organism contains more than 41000 kinds of small molecule metabolites, the metabolites play a very critical role in the process of the organism playing biological functions, the small molecule metabolites in the organism are characterized, the distribution and change characteristics of the small molecule metabolites are mastered, and the functions and mechanisms of the metabolites in the life activities can be deeply understood.
MALDI-MS (Matrix-assisted Laser Desorption Ionization Mass Spectrometry) technology plays an important role in the analysis of components such as lipids, proteins and polypeptides. However, MALDI-MS has been faced with a very important problem: the analysis of different chemical components depends on the selection of a matrix to a great extent, and the traditional matrixes such as Sinapic Acid (SA), 2, 5-dihydroxybenzoic acid (DHB), alpha-nitrile-4-hydroxycinnamic acid (CHCA) and the like are easy to generate phenomena such as fragmentation, intermolecular association and the like in the MALDI analysis process, so that serious background interference is generated in the range of m/z <1000, and the analysis of small molecule metabolites is seriously influenced.
In recent years, in order to overcome the problem of background interference of the matrix, researchers have developed novel matrix materials such as organic small molecules (e.g., 9 aminoacridine, proton sponge), organic salts (e.g., naphthyl ethylenediamine hydrochloride, 1, 5-naphthalene diamine hydrochloride), nanomaterials (e.g., carbon nanotubes, graphene), and the like for analysis of small molecule metabolites. These matrices have low background interference and present certain advantages in the analysis of small molecule metabolites. However, the preparation process of the nano material is complex and expensive, and a plurality of isotope peaks can appear in the organic salt matrix in the analysis process to interfere with the determination of other small molecule metabolites. In addition, these matrices have a problem of insufficient sensitivity when analyzing low levels of functional metabolites.
Disclosure of Invention
In order to improve the detection sensitivity and realize high coverage analysis of small molecule metabolites, the invention aims to provide a MALDI-MS method for detecting small molecule metabolites by using 1,1 '-binaphthyl-2, 2' -diamine as a matrix and application thereof. The invention takes 1,1 '-binaphthyl-2, 2' -diamine as a matrix, has the characteristics of low background interference and capability of obviously improving the detection sensitivity of MALDI-MS analysis of micromolecular metabolites.
The first object of the present invention: provides a MALDI-MS method for detecting small molecule metabolites by using 1,1 '-binaphthyl-2, 2' -diamine as a matrix.
The second object of the present invention: provides the application of 1,1 '-binaphthyl-2, 2' -diamine as MALDI-MS matrix.
In order to realize the purpose, the invention discloses the following technical scheme:
first, the present invention discloses the use of 1,1 '-binaphthyl-2, 2' -diamine as a MALDI-MS matrix, for example, for the detection, analysis, etc., of small molecule metabolites in animal tissues and plant tissues.
As a further technical solution, the small molecule metabolite includes: cholines (e.g., choline, glycerophosphocholine, etc.), carnitines (e.g., carnitine, C2:0 carnitine, etc.), polyamines (e.g., spermine, spermidine, etc.), organic acids (e.g., taurine, succinic acid, etc.), amino acids (e.g., arginine, glutamic acid, etc.), nucleosides (e.g., inosine, uridine, etc.), nucleotides (e.g., adenylic acid, inosinic acid, etc.), nitrogenous bases (e.g., xanthine, hypoxanthine, etc.), any one or more of fatty acids (e.g., linoleic acid, arachidonic acid, etc.), cholesterol (e.g., cholesterol sulfate, etc.), flavones (e.g., baicalein, wogonin, etc.), tanshinones (e.g., tanshinone I, tanshinone IIA, etc.), phenolic acids (e.g., caffeic acid, ferulic acid, etc.), polysaccharides (e.g., phytodisaccharides, phytotrisaccharides, etc.), polypeptides (e.g., glutathione, etc.), phospholipids (e.g., phosphatidylcholine, phosphatidylethanolamine, etc.).
Secondly, the invention discloses a MALDI-MS method for detecting micromolecular metabolites by taking 1,1 '-binaphthyl-2, 2' -diamine as a matrix, which comprises the following steps:
(1) transferring the frozen section made of the fresh biological tissue onto a conductive glass slide, and performing vacuum pumping treatment to obtain a tissue section;
(2) taking 1,1 '-binaphthyl-2, 2' -diamine as a matrix, preparing the matrix and a solvent into a solution, placing the solution on the tissue slice in the step (1), and performing MALDI mass spectrometry; and then carrying out targeted MALDI mass spectrum data extraction on different kinds of micromolecule metabolites in the biological tissue to obtain the ion intensity value of the micromolecule metabolites, thus obtaining the target MALDI mass spectrum.
As a further technical scheme, in the step (1), the fresh biological tissue comprises animal tissue and plant tissue.
As a further technical scheme, the thickness of the animal tissue section is 10-18 microns (preferably 12 mm). The slice thickness is too thick, so that the slice is easily interfered by non-target materials in the MALDI-MS analysis process, and the ionization efficiency of small molecule metabolites is influenced; if the thickness of the slice is too thin, the tissue is easy to shrink, so that the accurate analysis of the mass spectrum on the tissue in situ can not be realized.
As a further technical scheme, the thickness of the plant tissue slice is 6-12 microns (preferably 8 mm). If the thickness of the slice is too thin, the tissue is easy to shrink, so that the accurate analysis of the mass spectrum on the tissue in situ can not be realized. Since plant tissues have cell walls, if the thickness of the slices is too thick, intracellular components are not easily detected and are easily interfered by non-target substances.
As a further technical scheme, in the step (1), the conductive glass slide is an ITO-indium tin oxide conductive glass slide, and the diagonal resistance value is 20-300 omega (preferably 40 omega).
As a further technical scheme, in the step (2), the solvent is an acetonitrile-water mixed solution containing trifluoroacetic acid. Preferably, the mass concentration of the trifluoroacetic acid is 0.1-0.3%.
As a further technical scheme, the volume ratio of acetonitrile to water in the acetonitrile-water mixed solution is 7: 3.
as a further technical scheme, in the step (2), the concentration of the matrix in the solution is 0.2-1 mg/mL; preferably 0.5 mg/mL.
As a further technical solution, in the step (2), the method for spraying the solution onto the tissue slices comprises: the spraying speed is 0.05-0.15 mL/min, the spraying temperature is 40-90 ℃, the distance between nozzle tracks is 2-5mm, and the spraying is circulated for 6-12 times. Preferably, the spraying speed is 0.10 mL/min, the spraying temperature is 80 ℃, the distance between the nozzle tracks is 3mm, and the spraying is performed for 10 times in a circulating manner. The invention finally determines the matrix and the matrix spraying condition through continuous exploration, and is beneficial to improving the sensitivity and the coverage of MALDI-MS for detecting small molecule metabolites.
Compared with the prior art, the invention has the following beneficial effects:
(1) the invention discovers that MALDI-MS analysis of biological tissues by taking 1,1 '-binaphthyl-2, 2' -diamine as a matrix not only has low background interference, but also obviously improves the detection sensitivity of small molecule metabolites. Therefore, the detection sensitivity of the small molecule metabolite can be improved, and the detection sensitivity is related to that 1,1 '-binaphthyl-2, 2' -diamine has stronger ultraviolet absorption at the emission wavelength of the MALDI laser of 355nm and has a large conjugated system.
(2) The invention discovers that 1,1 '-binaphthyl-2, 2' -diamine is used as a substrate, high sensitivity detection on small molecular substances is realized, and simultaneously high coverage analysis on a plurality of metabolites in biological tissues can be realized, wherein the detected metabolite types comprise: choline, carnitine, polyamine, organic acid, amino acid, nucleoside, nucleotide, nitrogenous base, fatty acid, cholesterol, flavone, tanshinone, phenolic acid, polysaccharide, polypeptide and phospholipid.
(3) When the matrix is used for detection, the method is simple and convenient to operate, high in sensitivity, good in detection repeatability of the small molecule metabolite and good in practical application value, and the relative deviation (RSD) of mass spectrum response is 4.35% -9.45%.
Drawings
The accompanying drawings, which are incorporated in and constitute a part of this specification, are included to provide a further understanding of the invention, and are incorporated in and constitute a part of this specification, illustrate exemplary embodiments of the invention and together with the description serve to explain the invention and not to limit the invention.
FIG. 1 is a structural formula of 1,1 '-binaphthyl-2, 2' -diamine and a UV absorption diagram thereof.
FIG. 2 is a MALDI-MS mass spectrum of 1,1 '-binaphthyl-2, 2' -diamine in positive and negative ion detection mode.
FIG. 3 shows the results of MALDI-MS mass spectrometry of representative small molecule metabolites in brain tissue in example 1.
FIG. 4 is the MALDI-MS mass spectrometry detection result of representative small molecule metabolites in Salvia miltiorrhiza Bunge var.
Detailed Description
It is to be understood that the following detailed description is exemplary and is intended to provide further explanation of the invention as claimed. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of exemplary embodiments according to the invention. As used herein, the singular forms also are intended to include the plural forms as well, unless the context clearly indicates otherwise, and it should be further understood that when the terms "comprises" and/or "comprising" are used in this specification, they specify the presence of the stated features, steps, operations, devices, components, and/or combinations thereof.
As described above, when MALDI-MS is used for analyzing small molecule metabolites using the conventional matrix, problems such as insufficient sensitivity and large background interference are caused, which makes it difficult to perform simultaneous high-coverage analysis of a plurality of small molecule metabolites in a biological sample. Therefore, the invention provides a MALDI-MS method for detecting small molecule metabolites by taking 1,1 '-binaphthyl-2, 2' -diamine as a matrix and application thereof; the invention will now be further described with reference to the accompanying drawings and detailed description.
Example 1
A MALDI-MS method for detecting small molecule metabolites by using 1,1 '-binaphthyl-2, 2' -diamine as a matrix comprises the following steps:
(1) taking fresh rat brain tissue, and preparing frozen sections of the rat brain with the thickness of 12 microns by adopting a Thermo CryoStar NX50NOVPD microtome, wherein 9 sections are counted;
(2) transferring the frozen brain tissue slices to an ITO-indium tin oxide conductive glass slide with the size of 25mm x 75mm and the diagonal resistance of about 40 omega by using a fiber brush, then placing the obtained brain tissue slices in a vacuum drier, and drying for 15 minutes under the vacuum condition;
(3) precisely weighing 5mg of 1,1 '-binaphthyl-2, 2' -diamine in a 10mL volumetric flask, adding 10mL of acetonitrile-water (7:3, v/v) solution containing trifluoroacetic acid, uniformly mixing by vortex, and performing ultrasonic treatment for 10 minutes to obtain a 1,1 '-binaphthyl-2, 2' -diamine matrix solution for later use, wherein the mass fraction of the trifluoroacetic acid is 0.1%;
(4) accurately weighing 100mg of 9 aminoacridine, placing the 9 aminoacridine in a 10mL volumetric flask, adding 10mL of acetonitrile-water (7:3, v/v) solution containing trifluoroacetic acid, uniformly mixing by vortex, and carrying out ultrasonic treatment for 10 minutes to obtain a 9 aminoacridine matrix solution for later use, wherein the mass fraction of the trifluoroacetic acid is 0.1%;
(5) precisely weighing 200mg of 2, 5-dihydroxybenzoic acid, placing in a 10mL volumetric flask, adding 10mL of acetonitrile-water (7:3, v/v) solution containing trifluoroacetic acid, uniformly mixing by vortex, and performing ultrasonic treatment for 10 minutes to obtain a 2, 5-dihydroxybenzoic acid matrix solution for later use, wherein the mass fraction of the trifluoroacetic acid is 0.1%;
(6) using HTX TM-SprayerTMRespectively spraying the 1,1 '-binaphthyl-2, 2' -diamine, 9 aminoacridine and 2, 5-dihydroxybenzoic acid matrix solutions onto 3 adjacent brain tissue slices by using a matrix spraying instrument, wherein the spraying conditions are as follows: the spraying rate is 0.10 mL/min, the spraying temperature is 80 ℃, the space between the nozzle tracks is 3mm, and the cycle time is 10 times;
(7) using a BrookRapiflex-MALDI-tissuetyperTM-TOF type mass spectrometry imaging system for mass spectrometry analysis of 3 brain tissue sections sprayed with 1,1 '-binaphthyl-2, 2' -diamine, 9 aminoacridine, 2, 5-dihydroxybenzoic acid matrix solution;
(8) using HTX TM-SprayerTMThe matrix spraying instrument sprays 1,1 '-binaphthyl-2, 2' -diamine matrix solution on 6 adjacent brain tissue slices, and the spraying conditions are set as follows: the spraying rate is 0.10 mL/min, the spraying temperature is 80 ℃, the space between the nozzle tracks is 3mm, and the cycle time is 10 times;
(9) using a BrookRapiflex-MALDI-tissuetyperTMThe TOF type mass spectrometry imaging system carries out mass spectrometry on 6 brain tissue slices sprayed with the 1,1 '-binaphthyl-2, 2' -diamine matrix solution;
(10) targeted mass spectrum data extraction is carried out on micromolecule metabolites in brain tissue slices through BrookFleximaging 5.0 data processing software, and ion intensity values of the micromolecule metabolites are obtained.
(11) Comparing the mass spectrum response intensity of representative small molecule metabolites in brain tissue slices when 1,1 '-binaphthyl-2, 2' -diamine, 9 aminoacridine, 2, 5-dihydroxybenzoic acid were used as matrices, the results are shown in table 1 below, and it can be seen that the detection sensitivity of small molecule metabolites can be significantly improved when 1,1 '-binaphthyl-2, 2' -diamine was used as a matrix.
TABLE 1
(11) When 1,1 '-binaphthyl-2, 2' -diamine is used as a matrix, the mass spectrum response intensity of representative small molecule metabolites in 6 adjacent brain tissue slices is compared, and the relative deviation (RSD) is calculated, and as a result, as shown in table 2 below, it can be seen that the detection reproducibility of small molecule metabolites is good and the relative deviation (RSD) of mass spectrum response is between 4.35% and 9.45% when 1,1 '-binaphthyl-2, 2' -diamine is used as a matrix.
TABLE 2
Example 2
A MALDI-MS method for detecting small molecule metabolites by using 1,1 '-binaphthyl-2, 2' -diamine as a matrix comprises the following steps:
(1) taking fresh radix Salviae Miltiorrhizae, and preparing frozen slice of radix Salviae Miltiorrhizae with 8 μm thickness by using Thermo CryoStar NX50NOVPD microtome;
(2) transferring the frozen salvia miltiorrhiza slices to an ITO-indium tin oxide conductive glass slide with the size of 25mm by 75mm and the diagonal resistance of about 40 omega by using a fiber brush; then placing the obtained salvia miltiorrhiza slices in a vacuum drier, and pumping for 15 minutes under the vacuum condition;
(3) precisely weighing 5mg of 1,1 '-binaphthyl-2, 2' -diamine in a 10mL volumetric flask, adding 10mL of acetonitrile-water (7:3, v/v) solution containing trifluoroacetic acid, uniformly mixing by vortex, and performing ultrasonic treatment for 10 minutes to obtain a matrix solution for later use, wherein the mass fraction of the trifluoroacetic acid is 0.1%;
(4) using HTX TM-SprayerTMCarrying out matrix spraying on the salvia miltiorrhiza slices by using a matrix spraying instrument, and setting the spraying conditions as follows: the spraying rate is 0.10 mL/min, the spraying temperature is 80 ℃, the space between the nozzle tracks is 3mm, and the cycle time is 10 times;
(5) using a BrookRapiflex-MALDI-tissuetyperTMThe TOF type mass spectrometry imaging system carries out mass spectrometry on the salvia miltiorrhiza tissue slices sprayed with the 1,1 '-binaphthyl-2, 2' -diamine matrix;
(6) and performing targeted mass spectrum data extraction on micromolecular metabolites in the salvia miltiorrhiza tissue slices through BrookFleximaging 5.0 data processing software to obtain the ionic strength value of the micromolecular metabolites.
Example 3
A MALDI-MS method for detecting small molecule metabolites by using 1,1 '-binaphthyl-2, 2' -diamine as a matrix comprises the following steps:
(1) taking fresh rat liver tissue, and preparing frozen sections of the rat liver tissue with the thickness of 10 and 18 microns by adopting a ThermoCryoStar NX50NOVPD slicer;
(2) transferring the frozen liver tissue slices to an ITO-indium tin oxide conductive glass slide with the size of 25mm x 75mm and the diagonal resistance of about 20 omega by using a fiber brush, then placing the obtained brain tissue slices in a vacuum drier, and drying for 15 minutes under the vacuum condition;
(3) precisely weighing 2mg of 1,1 '-binaphthyl-2, 2' -diamine in a 10mL volumetric flask, adding 10mL of acetonitrile-water (7:3, v/v) solution containing trifluoroacetic acid, uniformly mixing by vortex, and performing ultrasonic treatment for 10 minutes to obtain 0.2mg/mL of 1,1 '-binaphthyl-2, 2' -diamine matrix solution for later use, wherein the mass fraction of the trifluoroacetic acid is 0.1%;
(4) using HTX TM-SprayerTMThe matrix spraying instrument is used for respectively spraying 0.2mg/mL of 1,1 '-binaphthyl-2, 2' -diamine matrix solution on the liver tissue slices, and the spraying conditions are set as follows: the spraying rate is 0.05 mL/min, the spraying temperature is 40 ℃, the space between the nozzle tracks is 3mm, and the cycle time is 10 times;
(5) using a BrookRapiflex-MALDI-tissuetyperTMThe TOF type mass spectrometry imaging system carries out mass spectrometry on the liver tissue section sprayed with the 1,1 '-binaphthyl-2, 2' -diamine matrix solution;
(6) targeted mass spectrum data extraction is carried out on micromolecule metabolites in liver tissue slices through BrookFleximaging 5.0 data processing software, and ion intensity values of the micromolecule metabolites are obtained.
Example 4
A MALDI-MS method for detecting small molecule metabolites by using 1,1 '-binaphthyl-2, 2' -diamine as a matrix comprises the following steps:
(1) taking fresh plant astragalus membranaceus tissue, and preparing frozen sections of the astragalus membranaceus tissue with the thickness of 6-12 microns by adopting a ThermoCryoStar NX50NOVPD slicer;
(2) transferring the frozen astragalus membranaceus tissue slices onto an ITO-indium tin oxide conductive glass slide with the size of 25mm x 75mm and the diagonal resistance of about 300 omega by using a fiber brush, then placing the obtained brain tissue slices into a vacuum drier, and drying for 15 minutes under a vacuum condition;
(3) precisely weighing 10mg of 1,1 '-binaphthyl-2, 2' -diamine in a 10mL volumetric flask, adding 10mL of acetonitrile-water (7:3, v/v) solution containing trifluoroacetic acid, uniformly mixing by vortex, and performing ultrasonic treatment for 10 minutes to obtain 1.0mg/mL of 1,1 '-binaphthyl-2, 2' -diamine matrix solution for later use, wherein the mass fraction of the trifluoroacetic acid is 0.3%;
(4) using HTX TM-SprayerTMThe matrix spraying instrument is used for respectively spraying 1.0mg/mL of 1,1 '-binaphthyl-2, 2' -diamine matrix solution on the astragalus membranaceus tissue slices, and the spraying conditions are set as follows: the spraying rate is 0.15 mL/min, the spraying temperature is 90 ℃, the space between the nozzle tracks is 3mm, and the cycle time is 10 times;
(5) using a BrookRapiflex-MALDI-tissuetyperTMThe TOF type mass spectrometry imaging system carries out mass spectrometry on the astragalus membranaceus tissue slices sprayed with the 1,1 '-binaphthyl-2, 2' -diamine matrix solution;
(6) targeted mass spectrum data extraction is carried out on micromolecule metabolites in the astragalus membranaceus tissue slices through BrookFleximaging 5.0 data processing software, and ion intensity values of the micromolecule metabolites are obtained.
And (3) detection results:
FIG. 1 is a diagram of the structural formula of 1,1 '-binaphthyl-2, 2' -diamine and its UV absorption, as can be seen: the 1,1 '-binaphthyl-2, 2' -diamine has stronger ultraviolet absorption at 355nm of MALDI laser, and the 1,1 '-binaphthyl-2, 2' -diamine has a large conjugated system in the structure.
FIG. 2 is a MALDI-MS mass spectrum of 1,1 '-binaphthyl-2, 2' -diamine in positive and negative ion detection mode. As can be seen from the figure: when 1,1 '-binaphthyl-2, 2' -diamine is used as a substrate, background interference is low.
FIG. 3 shows the MALDI-MS mass spectrometry detection results of representative small molecule metabolites in brain tissue. As can be seen from the figure: small molecule metabolites detected in brain tissue sections include amino acids, nitrogenous bases, nucleotides, cholesterol, fatty acids, choline, carnitine, creatine, phospholipids, and the like.
FIG. 4 shows the MALDI-MS mass spectrum detection result of representative small molecule metabolites in Salvia miltiorrhiza. As can be seen from the figure: the small molecule metabolites detected from the tissue slices of Salvia miltiorrhiza include tanshinol, salvianolic acid, tanshinone, choline, and tanshinol.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (30)
1. A MALDI-MS method for detecting small molecule metabolites by using 1,1 '-binaphthyl-2, 2' -diamine as a matrix is characterized by comprising the following steps:
(1) transferring the frozen section made of the fresh biological tissue onto a conductive glass slide, and performing vacuum pumping treatment to obtain a tissue section;
(2) taking 1,1 '-binaphthyl-2, 2' -diamine as a matrix, preparing the matrix and a solvent into a solution, placing the solution on the tissue slice in the step (1), and performing MALDI mass spectrometry; then, carrying out targeted MALDI mass spectrum data extraction on different kinds of micromolecule metabolites in biological tissues to obtain ion intensity values of the micromolecule metabolites;
the small molecule metabolites include: choline, carnosine, polyamine, organic acid, amino acid, nucleoside, nucleotide, nitrogenous base, fatty acid, cholesterol, flavone, tanshinone, phenolic acid, polysaccharide, polypeptide, and phospholipid.
2. The method of claim 1, wherein the choline comprises glycerophosphocholine.
3. The method of claim 1, wherein the carnitine is C2:0 carnitine.
4. The method of claim 1, wherein the polyamines are spermine and spermidine.
5. The method of claim 1, wherein the organic acid is taurine or succinic acid.
6. The method of claim 1, wherein the amino acids are arginine, glutamic acid.
7. The method of claim 1, wherein the nucleoside is inosine or uridine.
8. The method of claim 1, wherein the nucleotide is adenylic acid or inosinic acid.
9. The method of claim 1, wherein the nitrogenous base is xanthine, hypoxanthine.
10. The method of claim 1, wherein the fatty acid is linoleic acid, arachidonic acid.
11. The method of claim 1, wherein the cholesterol is cholesterol sulfate.
12. The method of claim 1, wherein the flavone is baicalein, wogonin.
13. The method as set forth in claim 1, wherein the tanshinone is tanshinone I or tanshinone IIA.
14. The method of claim 1, wherein the phenolic acid is caffeic acid, ferulic acid.
15. The method of claim 1, wherein the polysaccharide is a plant disaccharide, a plant trisaccharide.
16. The method of claim 1, wherein the polypeptide is glutathione.
17. The method of claim 1, wherein the phospholipid is phosphatidylcholine, phosphatidylethanolamine.
18. The method of claim 1, wherein in step (1), the fresh biological tissue comprises animal tissue and plant tissue.
19. The method of claim 18, wherein the thickness of the animal tissue section is 10-18 microns.
20. The method of claim 19, wherein the thickness of the animal tissue section is 12 microns.
21. The method of claim 18, wherein the plant tissue slices have a thickness of 6 to 12 microns.
22. The method of claim 21, wherein the plant tissue slices have a thickness of 8 microns.
23. The method of claim 1, wherein in step (1), the conductive glass slide is an ITO-indium tin oxide conductive glass slide, and the diagonal resistance value is 20-300 Ω.
24. The method according to claim 1, wherein in the step (2), the solvent is an acetonitrile-water mixed solution containing trifluoroacetic acid.
25. The method of claim 24, wherein the trifluoroacetic acid is present at a mass concentration of 0.1 to 0.3%.
26. The method of claim 24, wherein the volume ratio of acetonitrile to water in the acetonitrile-water is 7: 3.
27. the method of claim 24, wherein in step (2), the concentration of the matrix in the solution is 0.2-1 mg/mL.
28. The method of claim 27, wherein the concentration of the matrix in the solution is 0.5 mg/mL.
29. The method of claim 1, wherein in step (2), the solution is applied to the tissue slices by: the spraying method is adopted, the spraying speed is 0.05-0.15 mL/min, the spraying temperature is 40-90 ℃, the distance between the nozzle tracks is 2-5mm, and the spraying is circulated for 6-12 times.
30. The method of claim 29, wherein the spraying is performed 10 cycles at a rate of 0.10 mL/min, a spraying temperature of 80 ℃, and a nozzle track spacing of 3 mm.
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| CN116930008B (en) * | 2023-09-18 | 2023-12-01 | 华东理工大学 | In-situ mass spectrum identification method for detecting organic matters on black carbon surface and application of in-situ mass spectrum identification method |
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| CN106814129A (en) * | 2015-11-30 | 2017-06-09 | 中国科学院化学研究所 | Purposes and Mass Spectrometry detection method of the hexagonal boron nitride nanosheet in matrix solution is prepared |
| CN107085033A (en) * | 2017-03-31 | 2017-08-22 | 中国科学院化学研究所 | N phenylnaphthalenes aminated compounds as MALDI matrix application |
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