CN110146586A - The MALDI-MS method and application of 1,1 '--2,2 '-diamines of dinaphthalene detection small molecule metabolites - Google Patents
The MALDI-MS method and application of 1,1 '--2,2 '-diamines of dinaphthalene detection small molecule metabolites Download PDFInfo
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Abstract
The MALDI-MS method and application of 1,1'- dinaphthalene -2,2'- diamines detection small molecule metabolites.The described method includes: 1) fresh bio tissue frozen section is transferred on conducting glass slides, carries out vacuum and drain processing, obtain histotomy；2) matrix and solvent are configured on the histotomy for being sprayed into step (1) after solution for matrix with 1,1'- dinaphthalene -2,2'- diamines, carry out the analysis of MALDI mass spectrum imaging；Then targeting MALDI mass spectrometric data is carried out to variety classes small molecule metabolites in biological tissue to extract, obtain its ion-intensity values.The present invention is that matrix carries out biological tissue MALDI-MS analysis with 1,1'- dinaphthalene -2,2'- diamines, not only background interference is low, and the detection sensitivity of small molecule metabolites is significantly improved, simultaneously, additionally it is possible to realize high covering analyzing while a variety of metabolins in biological tissue.
Description
Technical field
The invention belongs to mass spectrum detection fields, and in particular to 1,1 '-dinaphthalene -2,2 '-diamines are as MALDI-MS base
The application of matter.
Background technique
It is only intended to increase understanding of the overall background of the invention for information disclosed in background of invention, without
It is existing well known to persons skilled in the art so to be considered as recognizing or imply that information composition has become in any form
Technology.
Small molecule metabolites (m/z < 1000) are life entity metabolism and the coefficient product of external environment, compared to gene
And albumen, it can more directly, accurately reflect the state of biosystem.41000 are had more than researches show that organism includes
Kind small molecule metabolites, these metabolins play extremely critical effect during physical exertion biological function, to machine
Body small molecular metabolin is characterized, and is grasped its distribution and variation characteristic, metabolin can be understood to a deeper level in life
Effect and mechanism in activity.
MALDI-MS (substance assistant laser desorpted ionized mass spectrum, Matrix-assisted Laser Desorption
Ionization Mass Spectrometry) technology played in the analysis of the ingredients such as lipid, albumen and polypeptide it is important
Effect.However MALDI-MS is but faced with always an extremely important problem: largely to the analysis of different chemical composition
The upper selection for relying on matrix, traditional sinapic acid (SA), 2,5-dihydroxybenzoic acid (DHB), α-itrile group -4- hydroxycinnamic acid
(CHCA) etc. phenomena such as matrix are easy to happen fragmentation and Interpolymer Association during MALDI analysis, causes the model of m/z < 1000
The interior serious background interference of generation is enclosed, the analysis of small molecule metabolites is seriously affected.
In recent years, in order to overcome the problems, such as that substrate background interferes, researchers develop small organic molecule (e.g., 9 ammonia in succession
Base acridine, proton sponge), organic salt (e.g., hydrochloride naphthodiamide, 1,5- naphthylenediamine hydrochloride), nano material (e.g., carbon nanometer
Pipe, graphene) etc. New-type matrix materials be used for small molecule metabolites analysis.The interference of these substrate backgrounds is low, in small molecule generation
Thank in the analysis of object that there is certain advantages.However nano material preparation process is complicated, and expensive, organic base status exists
Will appear many isotopic peaks in analytic process interferes other small molecule metabolites to measure.In addition, these matrix are analyzing low contain
It there is under-sensitive when the functional metabolism object of amount.
Summary of the invention
In view of the above problems, in order to improve detection sensitivity, the high covering analyzing of small molecule metabolites is realized, originally
Invention is intended to provide the MALDI-MS method and application that small molecule metabolites are detected using 1,1 '--2,2 '-diamines of dinaphthalene as matrix.
For the present invention with 1,1 '-dinaphthalene -2,2 '-diamines are matrix, have background interference low, can significantly improve MALDI-MS and analyze small point
The characteristics of detection sensitivity of sub- metabolin.
An object of the present disclosure: it provides and detects small molecule metabolites by matrix of 1,1 '--2,2 '-diamines of dinaphthalene
MALDI-MS method.
Second purpose of the invention: 1,1 '-applications of -2,2 '-diamines of dinaphthalene as MALDI-MS matrix are provided.
For achieving the above object, the invention discloses following technical proposals:
Firstly, the present invention discloses 1,1 '-dinaphthalene -2, application of the 2 '-diamines as MALDI-MS matrix, e.g., to animal groups
It knits, detection, the analysis of plant tissue small molecular metabolin etc..
As further technical solution, the small molecule metabolites include: choline (e.g., choline, choline glycerophosphatide
Deng), carnitine class (e.g., carnitine, C2:0 carnitine etc.), polyamines class (e.g., spermine, spermidine etc.), organic acid (e.g., taurine, amber
Amber acid etc.), amino acids (e.g., arginine, glutamic acid etc.), nucleosides (e.g., inosine, uridine etc.), nucleotide (e.g., adenylate, flesh
Thuja acid etc.), nitrogenous base (e.g., xanthine, hypoxanthine etc.), fatty acid (e.g., linoleic acid, arachidonic acid etc.), cholesterol
(e.g., cholesterol sulfate etc.), flavones (e.g., baicalein, wogonin etc.), tanshinone (e.g., Tanshinone I, tanshinone IIA etc.),
Phenolic acid (e.g., caffeic acid, ferulic acid etc.), polysaccharide (e.g., plant disaccharides, plant trisaccharide etc.), polypeptide (e.g., glutathione etc.), phosphorus
Any one or more in rouge (e.g., phosphatidyl choline, phosphatidyl-ethanolamine etc.).
Secondly, the present invention discloses one kind with 1,1 '-dinaphthalene -2,2 '-diamines are that matrix detects small molecule metabolites
MALDI-MS method, includes the following steps:
(1) fresh bio tissue frozen section is transferred on conducting glass slides, carries out vacuum and drain processing, obtains group
Knit slice；
(2) with 1,1 '-dinaphthalene -2,2 '-diamines are matrix, and matrix and solvent are configured to solution and are placed on step (1)
On histotomy, the analysis of MALDI mass spectrum imaging is carried out；Then variety classes small molecule metabolites in biological tissue are targeted
MALDI mass spectrometric data extract, obtain its ion-intensity values to get.
As further technical solution, in step (1), the fresh bio tissue includes animal tissue and plant group
It knits.
As further technical solution, the animal tissue sections with a thickness of 10-18 microns (preferably 12 millimeters).It cuts
Piece thickness is blocked up, and interference by non-target is easy in MALDI-MS analytic process, and influence small molecule metabolites from
Sonization efficiency；Slice thickness is excessively thin, then easily causes tissue shrinkage, to cannot achieve accurate analysis of the mass spectrum to tissue in situ.
As further technical solution, the plant tissue slice with a thickness of 6-12 microns (preferably 8 millimeters).Slice
Thickness is excessively thin, then easily causes tissue shrinkage, to cannot achieve accurate analysis of the mass spectrum to tissue in situ.Since plant tissue has
Cell wall be easy to cause intracellular members to be not easy to be detected if slice thickness is blocked up, and the interference vulnerable to non-target.
As further technical solution, in step (1), the conducting glass slides are that ITO- tin indium oxide conduction carries glass
Piece, diagonal line resistance value are 20-300 Ω (preferably 40 Ω).
As further technical solution, in step (2), the solvent is the acetonitrile-water mixed solution containing trifluoroacetic acid.
Preferably, the mass concentration of the trifluoroacetic acid is 0.1-0.3%.
As further technical solution, the volume ratio of acetonitrile and water is 7:3 in the acetonitrile-water mixed solution.
As further technical solution, in step (2), the concentration of the solution mesostroma is 0.2-1mg/mL；It is preferred that
For 0.5mg/mL.
As further technical solution, in step (2), the method by solution spraying to histotomy are as follows: spray
Applying rate is 0.05-0.15mL/ minutes, and spraying temperature is 40-90 DEG C, and nozzle track spacing is 2-5mm, circulation spraying 6-12
It is secondary.Preferably, the spray rate is 0.10mL/ minutes, and spraying temperature is 80 DEG C, and nozzle track spacing is 3mm, circulation spraying
10 times.The selection of matrix and matrix coverage condition can all influence the mass spectral analysis quality of tissue target, and the present invention is not by
It is disconnected to explore, it finally determines above-mentioned matrix and matrix spraying conditions, helps to improve the spirit of MALDI-MS detection small molecule metabolites
Sensitivity and coverage.
Compared with prior art, the present invention achieve it is following the utility model has the advantages that
(1) it is a discovery of the invention that by with 1,1 '-dinaphthalene -2,2 '-diamines are that matrix carries out biological tissue MALDI-MS points
Analysis, not only background interference is low, but also significantly improves the detection sensitivity of small molecule metabolites.Why small molecule can be improved
The detection sensitivity of metabolin, and 1,1 '-dinaphthalene -2,2 '-diamines have stronger at the launch wavelength 355nm of MALDI laser
UV absorption, and the big conjugated system having of itself is related.
(2) it is a discovery of the invention that with 1,1 '-dinaphthalene -2,2 '-diamines are matrix, highly sensitive to small-molecule substance realizing
While degree detection, additionally it is possible to high covering analyzing while a variety of metabolins in biological tissue is realized, wherein the metabolism detected
Species include: choline, carnitine, polyamines, organic acid, amino acid, nucleosides, nucleotide, nitrogenous base, fatty acid, cholesterol, Huang
Ketone, tanshinone, phenolic acid, polysaccharide, polypeptide, phospholipid.
(3) when being detected using matrix of the invention, not only easy to operate, high sensitivity, but also small molecule metabolites
Detection favorable reproducibility, the relative deviation (RSD) of mass spectrum response between 4.35%-9.45%, therefore the present invention have it is good
Practical application value.
Detailed description of the invention
The Figure of description for constituting a part of the invention is used to provide further understanding of the present invention, and of the invention shows
Examples and descriptions thereof are used to explain the present invention for meaning property, does not constitute improper limitations of the present invention.
Fig. 1 is the structural formula and its UV absorption figure of 1,1 '--2,2 '-diamines of dinaphthalene.
Fig. 2 is MALDI-MS mass spectrogram of 1,1 '--2,2 '-diamines of dinaphthalene under positive and negative ion detection pattern.
Fig. 3 is the MALDI-MS Mass Spectrometer Method result of representative small molecule metabolites in 1 mesencephalic tissue of embodiment.
Fig. 4 is the MALDI-MS Mass Spectrometer Method result of representative small molecule metabolites in violet flower danshen in embodiment 2.
Specific embodiment
It is noted that following detailed description is all illustrative, it is intended to provide further instruction to the present invention.Unless another
It indicates, all technical and scientific terms used herein has usual with general technical staff of the technical field of the invention
The identical meanings of understanding.
It should be noted that term used herein above is merely to describe specific embodiment, and be not intended to restricted root
According to exemplary embodiments of the present invention.Such as, used herein, unless the context clearly indicates otherwise, otherwise singular shape
Formula is also intended to include plural form, additionally, it should be understood that, when in the present specification using term "comprising" and/or " packet
Include " when, indicate existing characteristics, step, operation, device, component and/or their combination.
As, it is described previously, due to there is spirit when carrying out MALDI-MS analysis small molecule metabolites using current matrix
The problem that sensitivity is insufficient, background interference is big causes to be difficult to realize height while a variety of small molecule metabolites in biological sample and covers
Analysis.Therefore, the present invention proposes that, with 1,1 '-dinaphthalene -2,2 '-diamines are the MALDI-MS method that matrix detects small molecule metabolites
And application；The present invention is further illustrated now in conjunction with the drawings and specific embodiments.
Embodiment 1
One kind is with 1,1 '-dinaphthalene -2, and 2 '-diamines are the MALDI-MS method that matrix detects small molecule metabolites, including such as
Lower step:
(1) fresh rat brain tissue is taken, makes 12 microns thick using Thermo CryoStar NX50NOVPD slicer
Mouse brain frozen section amounts to 9；
(2) brain tissue frozen section is transferred to size is 25mm*75mm and diagonal line resistance is about 40 with fiber hairbrush
On the ITO- tin indium oxide conducting glass slides of Ω, then obtained brain tissue slice is placed in vacuum desiccator, in vacuum item
It is drained under part 15 minutes；
(3) the 1 of precision weighing 5mg, 1 '-dinaphthalene -2,2 '-diamines are placed in 10mL volumetric flask, and 10mL second containing trifluoro is added
Acetonitrile-water (7:3, v/v) solution of acid is vortexed and mixes, and ultrasound 10 minutes obtains 1,1 '-dinaphthalene -2,2 '-diamines matrix solutions
Spare, the mass fraction of the trifluoroacetic acid is 0.1%；
(4) accurate 9 aminacrines for weighing 100mg are placed in 10mL volumetric flask, and acetonitrile-of the 10mL containing trifluoroacetic acid is added
Water (7:3, v/v) solution is vortexed and mixes, and ultrasound 10 minutes, it is spare to obtain 9 aminacrine matrix solutions, the trifluoroacetic acid
Mass fraction is 0.1%；
(5) the accurate 2,5-dihydroxybenzoic acid for weighing 200mg is placed in 10mL volumetric flask, and 10mL is added and contains trifluoroacetic acid
Acetonitrile-water (7:3, v/v) solution, be vortexed and mix, ultrasound 10 minutes, it is spare to obtain 2,5-dihydroxybenzoic acid matrix solution,
The mass fraction of the trifluoroacetic acid is 0.1%；
(6) HTX TM-Sprayer is usedTMMatrix sprays instrument respectively by above-mentioned -2,2 '-diamines of 1,1 '-dinaphthalene, 9 amino a word used for translations
Pyridine, 2,5-dihydroxybenzoic acid matrix solution spray on 3 adjacent brain tissue slices, set spraying conditions are as follows: spraying speed
Rate is 0.10mL/ minutes, and spraying temperature is 80 DEG C, and nozzle track spacing is 3mm, and cycle-index is 10 times；
(7) Brooker Rapiflex-MALDI-tissuetyper is utilizedTM- TOF type mass spectrum imaging system to having sprayed 1,
1 '--2,2 '-diamines of dinaphthalene, 9 aminacrines, 2,5- dihydroxybenzoic acid matrix solution 3 brain tissue slices carry out mass spectrum point
Analysis；
(8) HTX TM-Sprayer is usedTMMatrix sprays instrument and 1,1 '--2,2 '-diamines matrix solutions of dinaphthalene is sprayed to phase
On 6 adjacent brain tissue slices, spraying conditions are set are as follows: spray rate is 0.10mL/ minutes, and spraying temperature is 80 DEG C, nozzle
For interorbital away from for 3mm, cycle-index is 10 times；
(9) Brooker Rapiflex-MALDI-tissuetyper is utilizedTM- TOF type mass spectrum imaging system to having sprayed 1,
6 brain tissue slices of 1 '--2,2 '-diamines matrix solution of dinaphthalene are analyzed by mass spectrometry；
(10) by 5.0 data processing software of Brooker FlexImaging to brain tissue slice small molecular metabolin into
Row targeting mass spectrometric data is extracted, its ion-intensity values is obtained.
(11) comparing ought be respectively using 1,1 '--2,2 '-diamines of dinaphthalene, 9 aminacrines, 2,5- dihydroxy-benzoic acid as matrix
When, the mass spectrum response intensity of representative small molecule metabolites in brain tissue slice, as a result as shown in table 1 below, it is seen that when with 1,1 '-
Dinaphthalene -2,2 '-diamines can significantly improve the detection sensitivity of small molecule metabolites when being matrix.
Table 1
(11) small point representative in more adjacent 6 brain tissue slices when 2 '-diamines are matrix when with 1,1 '-dinaphthalene -2
The mass spectrum response intensity of sub- metabolin calculates relative deviation (RSD), as a result as shown in table 2 below, it is seen that when with 1,1 '-dinaphthalene -2,
The detection favorable reproducibility of small molecule metabolites when 2 '-diamines are matrix, the relative deviation (RSD) of mass spectrum response is in 4.35%-
Between 9.45%.
Table 2
Embodiment 2
One kind is with 1,1 '-dinaphthalene -2, and 2 '-diamines are the MALDI-MS method that matrix detects small molecule metabolites, including such as
Lower step:
(1) fresh plant red sage root is taken, makes 8 microns thick using Thermo CryoStar NX50NOVPD slicer
Radix Salviae Miltiorrhizae frozen section；
(2) Radix Salviae Miltiorrhizae frozen section is transferred to size is 25mm*75mm and diagonal line resistance is about 40 Ω with fiber hairbrush
ITO- tin indium oxide conducting glass slides on；Then obtained Radix Salviae Miltiorrhizae slice is placed in vacuum desiccator, under vacuum conditions
It drains 15 minutes；
(3) the 1 of precision weighing 5mg, 1 '-dinaphthalene -2,2 '-diamines are placed in 10mL volumetric flask, and 10mL second containing trifluoro is added
Acetonitrile-water (7:3, v/v) solution of acid is vortexed and mixes, and ultrasound 10 minutes obtains that matrix solution is spare, the trifluoroacetic acid
Mass fraction is 0.1%；
(4) HTX TM-Sprayer is usedTMMatrix sprays the matrix spraying that instrument carries out Radix Salviae Miltiorrhizae slice, sets spraying conditions
Are as follows: spray rate is 0.10mL/ minutes, and spraying temperature is 80 DEG C, and nozzle track spacing is 3mm, and cycle-index is 10 times；
(5) Brooker Rapiflex-MALDI-tissuetyper is utilizedTM- TOF type mass spectrum imaging system to having sprayed 1,
The Radix Salviae Miltiorrhizae histotomy of 1 '--2,2 '-diamines matrix of dinaphthalene is analyzed by mass spectrometry；
(6) by 5.0 data processing software of Brooker FlexImaging to Radix Salviae Miltiorrhizae histotomy small molecular metabolin
It carries out targeting mass spectrometric data to extract, obtains its ion-intensity values.
Embodiment 3
One kind is with 1,1 '-dinaphthalene -2, and 2 '-diamines are the MALDI-MS method that matrix detects small molecule metabolites, including such as
Lower step:
(1) fresh rat liver organization is taken, makes 10,18 microns using ThermoCryoStar NX50NOVPD slicer
Thick rat liver frozen section；
(2) liver organization frozen section is transferred to size is 25mm*75mm and diagonal line resistance is about with fiber hairbrush
On the ITO- tin indium oxide conducting glass slides of 20 Ω, then obtained brain tissue slice is placed in vacuum desiccator, in vacuum
Under the conditions of drain 15 minutes；
(3) the 1 of precision weighing 2mg, 1 '-dinaphthalene -2,2 '-diamines are placed in 10mL volumetric flask, and 10mL second containing trifluoro is added
Acetonitrile-water (7:3, v/v) solution of acid is vortexed and mixes, and ultrasound 10 minutes obtains the 1 of 0.2mg/mL, 1 '-dinaphthalene -2, and 2 '-two
Amido matter solution for standby, the mass fraction of the trifluoroacetic acid are 0.1%；
(4) HTX TM-Sprayer is usedTMMatrix sprays instrument respectively by -2,2 '-two amido of the 1,1 ' of 0.2mg/mL-dinaphthalene
On matter solution spraying to liver tissue slices, spraying conditions are set are as follows: spray rate is 0.05mL/ minutes, spraying temperature 40
DEG C, nozzle track spacing is 3mm, and cycle-index is 10 times；
(5) Brooker Rapiflex-MALDI-tissuetyper is utilizedTM- TOF type mass spectrum imaging system to having sprayed 1,
The hepatic tissue section of 1 '--2,2 '-diamines matrix solution of dinaphthalene is analyzed by mass spectrometry；
(6) by 5.0 data processing software of Brooker FlexImaging to liver tissue slices small molecular metabolin
It carries out targeting mass spectrometric data to extract, obtains its ion-intensity values.
Embodiment 4
One kind is with 1,1 '-dinaphthalene -2, and 2 '-diamines are the MALDI-MS method that matrix detects small molecule metabolites, including such as
Lower step:
(1) fresh plant callus is taken, makes 6,12 microns using ThermoCryoStar NX50NOVPD slicer
Thick callus frozen section；
(2) callus frozen section is transferred to size is 25mm*75mm and diagonal line resistance is about with fiber hairbrush
On the ITO- tin indium oxide conducting glass slides of 300 Ω, then obtained brain tissue slice is placed in vacuum desiccator, in vacuum
Under the conditions of drain 15 minutes；
(3) the 1 of precision weighing 10mg, 1 '-dinaphthalene -2,2 '-diamines are placed in 10mL volumetric flask, and 10mL second containing trifluoro is added
Acetonitrile-water (7:3, v/v) solution of acid is vortexed and mixes, and ultrasound 10 minutes obtains the 1 of 1.0mg/mL, 1 '-dinaphthalene -2, and 2 '-two
Amido matter solution for standby, the mass fraction of the trifluoroacetic acid are 0.3%；
(4) HTX TM-Sprayer is usedTMMatrix sprays instrument respectively by -2,2 '-two amido of the 1,1 ' of 1.0mg/mL-dinaphthalene
Matter solution spraying is sliced to callus, sets spraying conditions are as follows: spray rate is 0.15mL/ minutes, spraying temperature 90
DEG C, nozzle track spacing is 3mm, and cycle-index is 10 times；
(5) Brooker Rapiflex-MALDI-tissuetyper is utilizedTM- TOF type mass spectrum imaging system to having sprayed 1,
The callus slice of 1 '--2,2 '-diamines matrix solution of dinaphthalene is analyzed by mass spectrometry；
(6) small molecular metabolin is sliced to callus by 5.0 data processing software of Brooker FlexImaging
It carries out targeting mass spectrometric data to extract, obtains its ion-intensity values.
Testing result:
Fig. 1 is 1,1 '-dinaphthalene -2, and the structural formula and its UV absorption figure of 2 '-diamines, as can be seen from the figure: 1,1 '-joins
2,2 '-diamines of naphthalene-has provided stronger UV absorption, and 1,1 '-dinaphthalene -2,2 '-diamines itself in MALDI laser 355nm
There is big conjugated system in structure.
Fig. 2 is MALDI-MS mass spectrogram of 1,1 '--2,2 '-diamines of dinaphthalene under positive and negative ion detection pattern.From figure
It can be seen that 1 '-dinaphthalene -2, when 2 '-diamines are matrix, background interference is lower with 1.
The MALDI-MS Mass Spectrometer Method result of representative small molecule metabolites in the brain tissue that Fig. 3 is shown.It can be with from figure
Find out: the small molecule metabolites detected in brain tissue slice include amino acid, nitrogenous base, nucleotide, cholesterol, fat
Acid, choline, carnitine, creatine, phosphatide etc..
Fig. 4 is the MALDI-MS Mass Spectrometer Method result of representative small molecule metabolites in Radix Salviae Miltiorrhizae.As can be seen from the figure: from
The small molecule metabolites that Radix Salviae Miltiorrhizae histotomy detects include danshensu, danshinolic acid, tanshinone, choline, Radix Salviae Miltiorrhizae glycol etc..
The above description is only a preferred embodiment of the present invention, is not intended to restrict the invention, for those skilled in the art
For member, the invention may be variously modified and varied.All within the spirits and principles of the present invention, it is made it is any modification,
Equivalent replacement, improvement etc., should all be included in the protection scope of the present invention.
Claims (10)
- Application of the 1.1,1'- dinaphthalene -2,2'- diamines as matrix in substance assistant laser desorpted ionized Mass Spectrometer Method.
- Substance assistant laser desorpted electricity of the 2.1,1'- dinaphthalene -2,2'- diamines as matrix small molecule metabolites in biological tissues From the application in Mass Spectrometer Method.
- 3. the application as described in requiring 2 in right, which is characterized in that the biological tissue knits comprising animal tissue and/or plant Tissue.
- 4. the application as described in requiring 2 or 3 in right, which is characterized in that the small molecule metabolites include: choline (preferably For choline, choline glycerophosphatide), carnitine class (preferably carnitine, C2:0 carnitine), polyamines class (preferably spermine, spermidine), have Machine acids (preferably taurine, succinic acid), amino acids (preferably arginine, glutamic acid), nucleosides (preferably inosine, urine Glycosides), nucleotide (preferably adenylate, inosinicacid), nitrogenous base (preferably xanthine, hypoxanthine), fatty acid (preferably Linoleic acid, arachidonic acid), cholesterol (preferably cholesterol sulfate), flavones (preferably baicalein, wogonin), Radix Salviae Miltiorrhizae Ketone (preferably Tanshinone I, tanshinone IIA), phenolic acid (preferably caffeic acid, ferulic acid), polysaccharide (preferably plant disaccharides, plant Object trisaccharide), polypeptide (preferably glutathione), any one in phosphatide (preferably phosphatidyl choline, phosphatidyl-ethanolamine) Or it is a variety of.
- 5. a kind of MALDI-MS method for detecting small molecule metabolites for matrix with 1,1'- dinaphthalene -2,2'- diamines, feature exist In including the following steps:(1) fresh bio tissue frozen section is transferred on conducting glass slides, carries out vacuum and drain processing, obtains tissue and cut Piece；(2) matrix and solvent are configured to for matrix by the tissue that solution is placed on step (1) with 1,1'- dinaphthalene -2,2'- diamines On slice, the analysis of MALDI mass spectrum imaging is carried out；Then variety classes small molecule metabolites in biological tissue are targeted MALDI mass spectrometric data extract, obtain its ion-intensity values to get.
- 6. the MALDI-MS of small molecule metabolites is detected using 1,1'- dinaphthalene -2,2'- diamines as matrix as claimed in claim 5 Method, which is characterized in that in step (1), the fresh bio tissue includes animal tissue and plant tissue.
- 7. the MALDI-MS of small molecule metabolites is detected using 1,1'- dinaphthalene -2,2'- diamines as matrix as claimed in claim 6 Method, which is characterized in that the animal tissue sections with a thickness of 10-18 microns, more preferably 12 millimeters；Preferably, described Plant tissue slice with a thickness of 6-12 microns, more preferably 8 millimeters.
- 8. the MALDI-MS of small molecule metabolites is detected using 1,1'- dinaphthalene -2,2'- diamines as matrix as claimed in claim 5 Method, which is characterized in that in step (1), the conducting glass slides are ITO- tin indium oxide conducting glass slides, diagonal line resistance value For 20-300 Ω, preferably 40 Ω.
- 9. the MALDI-MS of small molecule metabolites is detected using 1,1'- dinaphthalene -2,2'- diamines as matrix as claimed in claim 5 Method, which is characterized in that in step (2), the solvent is the acetonitrile-water mixed solution containing trifluoroacetic acid；Preferably, described three The mass concentration of fluoroacetic acid is 0.1-0.3%；Preferably, the volume ratio of acetonitrile and water is 7:3 in the acetonitrile-water；Preferably, in step (2), the concentration of the solution mesostroma is 0.2-1mg/mL；Preferably 0.5mg/mL.
- 10. as claim 5-9 it is described in any item using 1,1'- dinaphthalene -2,2'- diamines as matrix detect small molecule metabolites MALDI-MS method, which is characterized in that in step (2), method solution being applied on histotomy are as follows: using spraying Method, spray rate are 0.05-0.15mL/ minutes, and spraying temperature is 40-90 DEG C, and nozzle track spacing is 2-5mm, circulation spray It applies 6-12 times；Preferably, the spray rate is 0.10mL/ minutes, and spraying temperature is 80 DEG C, and nozzle track spacing is 3mm, circulation spray It applies 10 times.
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| CN113030236A (en) * | 2019-12-09 | 2021-06-25 | 深圳先进技术研究院 | In-situ analysis method for lipid compounds in biological tissues |
| CN113588768A (en) * | 2021-05-18 | 2021-11-02 | 国家卫生健康委科学技术研究所 | Mass spectrometry method for quantifying endogenous metabolites in tissues in molecular image mode |
| CN116930008A (en) * | 2023-09-18 | 2023-10-24 | 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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| 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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