CN112986374A - Laser desorption ionization mass spectrum matrix, detection method, mass spectrum and application of mass spectrum in neurotransmitter detection - Google Patents

Abstract

The invention discloses a laser desorption ionization mass spectrometry matrix, a detection method, a mass spectrometry and application of the mass spectrometry in neurotransmitter detection, belongs to the technical field of mass spectrometry detection, and can solve the problems that the matrix in the prior art needs longer detection time when detecting neurotransmitter and cannot meet the clinical detection requirement on the neurotransmitter. The gold tris (triphenylphosphine) tetrafluoroborate is used as a matrix in the laser desorption ionization mass spectrum, and the gold tris (triphenylphosphine) tetrafluoroborate forms gold nanoclusters (Au NCLs) which can bear ultraviolet light in the derivation process, and the gold nanoclusters can absorb more ultraviolet light in the detection process of the laser desorption ionization mass spectrum, so that the ionization energy is increased, the ionization efficiency is improved, the detection rate is improved, the detection time is shortened, and the clinical detection requirement on the neurotransmitter can be met.

Description

Laser desorption ionization mass spectrum matrix, detection method, mass spectrum and application of mass spectrum in neurotransmitter detection

Technical Field

The invention relates to the technical field of mass spectrum detection, in particular to a laser desorption ionization mass spectrum matrix, a detection method, a mass spectrum and application of the mass spectrum in neurotransmitter detection.

Background

Neurotransmitters, which are messengers in synaptic signalling, are important chemical substances in the human brain. Any imbalance in neurotransmitters can cause serious psychiatric disorders such as parkinson's disease, schizophrenia, and alzheimer's disease. Therefore, monitoring the concentration of various neurotransmitters is highly desirable for the study and diagnosis of such mental disorders.

Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) technology is widely applied to detection of small molecules such as neurotransmitter and the like due to the advantages of flexibility, high sensitivity, compatibility, diversity of mass analyzers and the like. However, the development of a highly specific organic compound as a substrate for rapid, sensitive and selective detection of neurotransmitters has been a formidable challenge.

Currently, in the prior art, because the pyranoid salt has high sensitivity and detection selectivity on various neurotransmitters, the pyranoid salt is used as a matrix in matrix-assisted laser desorption ionization mass spectrometry for detecting the concentration of the neurotransmitter, but the reaction rate of derivatization of the pyranoid salt and the neurotransmitter is slow, the detection time is long, and the clinical requirement on detection of the neurotransmitter cannot be met.

Disclosure of Invention

In view of the above, the invention provides a laser desorption ionization mass spectrometry matrix, a detection method, a mass spectrometry and application of the mass spectrometry in neurotransmitter detection, and can solve the problems that the matrix and the neurotransmitter in the prior art are slow in derivatization reaction, long in detection time and incapable of meeting requirements for detecting the neurotransmitter clinically.

In order to achieve the purpose, the invention provides the following technical scheme:

a laser desorption ionization mass spectrometry matrix comprising gold tris (triphenylphosphine) tetrafluoroborate.

The invention also provides a mass spectrum applying the laser desorption ionization mass spectrum matrix, wherein the mass spectrum is a matrix-assisted laser desorption ionization mass spectrum; the matrix comprises gold tris (triphenylphosphine) tetrafluoroborate.

The invention also provides a detection method applying the laser desorption ionization mass spectrometry matrix, wherein the substance to be detected is selected from polypeptide containing primary amine groups;

the detection method uses matrix-assisted laser desorption ionization mass spectrometry; the matrix comprises gold tris (triphenylphosphine) tetrafluoroborate.

As a still further scheme of the invention: the molecular weight of the polypeptide containing the primary amine group is less than 500.

In the derivatization process of the invention, tris (triphenylphosphine) gold tetrafluoroborate ([ Ph ] in the presence of a polypeptide containing a primary amine group₃PAu]₃O⁺BF₄ ^-) The first gold-oxygen-gold bond of (a) is cleaved. After cleavage, a different ligand containing gold ions is released, e.g. [ Ph ]₃PAu]⁺、[(Ph₃P)₂Au]⁺And [ (Ph)₃PAu)₃]⁺. Then, the nitrogen atom in the primary amine group of the polypeptide containing the primary amine group serves as an aggregation center, and the ammonium derivative having a gold-nitrogen-gold bond is regenerated and gold nanoparticles are formed. However, only [ Ph₃PAu]⁺Participate in the formation of gold nanoparticles to [ (Ph)₃P)₂Au]⁺And [ (Ph)₃PAu)₃]⁺Do not participate because their gold ions have already been Ph₃The P ligand is stable. There have been a number of research efforts regarding the mechanism by which organophosphines support gold nanoparticles around the nitrogen atom of a primary amine. It is reported that one central nucleolar atom can carry up to six gold cluster units. The metallophilic interaction can be achieved under two conditions, where the gold atom should be a single atom, and the distance between the two gold atoms is as close as possible in a linear orientation. The metallophilic phase interacts much weaker than the ionic and covalent bonds, but it provides strength equivalent to hydrogen bonding. As shown in fig. 1, is a detailed mechanism of the formation of gold nanoparticles such as monomers, dimers, and trimers. In the presence of catechol groups in the polypeptide, the gold nanoclusters may be further converted to gold nanoparticles. As shown in fig. 2, the catechol group releases one or two electrons during oxidation, and the released electrons can be used to reduce the gold ions of the gold nanoparticles to neutral gold nanoparticles. However, non-catecholamines such as gamma-aminobutyric acid and histamine cannot form gold nanoparticles, and they can only generate gold nanoclusters.

As a still further scheme of the invention: the substance to be detected is at least one selected from dopamine, norepinephrine, 5-hydroxytryptamine, gamma-aminobutyric acid, histamine and tyramine.

As a still further scheme of the invention: the method comprises at least the following steps:

dissolving a tris (triphenylphosphine) gold tetrafluoroborate matrix and a substance to be detected in an organic solvent respectively to form a matrix solution and a solution to be detected;

and mixing the matrix solution and the solution to be detected to form a mixed solution, spotting the mixed solution on a matrix-assisted laser desorption ionization target plate, and analyzing by using the matrix-assisted laser desorption ionization mass spectrometry after drying.

As a still further scheme of the invention: the organic solvent is at least one selected from methanol, acetone, isopropanol and acetonitrile.

Preferably, the organic solvent is acetonitrile.

As shown in FIG. 3, the present invention compares the abundance values of the final product obtained by 4 different solvents, namely Acetone (ACE), Acetonitrile (ACN), Isopropanol (IPA) and Methanol (MA), and the results show that the abundance of acetonitrile is superior to that of other solvents.

As a still further scheme of the invention: in the mixed solution, the molar ratio of the tris (triphenylphosphine) gold tetrafluoroborate to the sample to be detected is more than 10: 1.

preferably, the molar ratio of the tris (triphenylphosphine) gold tetrafluoroborate to the sample to be tested is independently selected from: 80:1, 70:1, 60:1, 50:1, 40:1, 30:1, 20:1, 10: 1.

As a still further scheme of the invention: the method comprises at least the following steps:

dissolving a tris (triphenylphosphine) gold tetrafluoroborate matrix and a substance to be detected in an organic solvent respectively to form a matrix solution and a solution to be detected;

mixing the matrix solution and the sample solution, and centrifuging to obtain a supernatant;

and (3) spotting the supernatant on a matrix-assisted laser desorption ionization target plate, and performing mass spectrometry after drying.

In the process of research and development, the invention is used for [ Ph₃PAu]₃O⁺BF₄ ^-Concentration of matrix and Standard additional matrixThe optimization was performed and the results are shown in fig. 4 and 5.

As shown in FIG. 4, is [ Ph₃PAu]₃O⁺At different concentrations, [ Ph ] is obtained₃PAu]⁺、[Ph₃PAu]₂N⁺R、[Ph₃PAu]₃N⁺The results show that when [ Ph ] is₃PAu]₃O⁺At a concentration of 100. mu. mol/L, [ Ph ]₃PAu]⁺、[Ph₃PAu]₂N⁺R、[Ph₃PAu]₃N⁺The abundance value of R is the best.

As shown in FIG. 5, at [ Ph₃PAu]₃O⁺And a neurotransmitter, wherein the standard additional matrix is alpha-cyano-4-hydroxycinnamic acid (CHCA), 2, 5-dihydroxybenzoic acid (DHB) or Sinapic Acid (SA), respectively, only CHCA does not react with the neurotransmitter, but only Ph is increased as shown in FIG. 5₃PAu]₃O⁺And signal strength of neurotransmitter products.

The invention also provides an application of the mass spectrum in neurotransmitter detection.

The beneficial effects of the invention include but are not limited to:

(1) the invention adopts tri (triphenylphosphine) gold tetrafluoroborate as a matrix in matrix-assisted laser desorption ionization mass spectrometry, and the tri (triphenylphosphine) gold tetrafluoroborate ([ Ph ] is₃PAu]₃O⁺BF₄ ^-) Gold nanoclusters (Au NCLs) which can bear ultraviolet light are formed in the process of derivatization of the gold nanoclusters with polypeptides containing primary amine groups, the gold nanoclusters can absorb more ultraviolet light in the process of MALDI-TOF-MS detection, so that ionization energy is increased, ionization efficiency is improved, detection rate is improved, detection time is shortened, the detection speed is about 10 times faster than that of the existing pyranoid salt serving as a matrix, and the detection requirement on neurotransmitter in clinic can be met.

(2) Qualitative and quantitative analysis of the neurotransmitters of the present invention is achieved by measuring the derivative products. Compared with other technologies, the existing mass spectrum coupling technology has the advantages of simplicity, high efficiency and the like. And isThe methods of the invention also reduce matrix interference, salt interference, and endogenous impurities. [ Ph ] used in the invention₃PAu]₃O⁺BF₄ ^-The matrix has high sensitivity, selectivity, rapid response time, molar absorption coefficient and derivatization efficiency.

(3) [ Ph ] of the invention₃PAu]₃O⁺BF₄ ^-The method can also derive various neurotransmitters simultaneously, can detect various neurotransmitters simultaneously, and has the advantages of high detection speed and high accuracy, and the accuracy and the detection speed of clinical detection are greatly improved.

Drawings

FIG. 1 shows the present invention [ Ph₃PAu]₃O⁺BF₄ ^-And the formation mechanism of gold monomers, dimers and trimers;

FIG. 2 shows [ Ph ] of the present invention₃PAu]⁺Schematic representation of the derived reactions with various neurotransmitters;

FIG. 3 is a graph of [ Ph ] in 100. mu. mol/L DA in combination with 100. mu. mol/L of a different solvent₃PAu]₃O⁺BF₄ ^-Upon derivatization reaction, [ Ph ] is formed₃PAu]⁺、[Ph₃PAu]₂N⁺R and [ Ph ]₃PAu]₃N⁺A schematic representation of the relative abundance of R products;

FIG. 4 shows (a) [ Ph ] at 10. mu. mol/L dopamine as neurotransmitter in the present invention₃PAu]₃O⁺BF₄ ^-And [ Ph₃PAu]⁺(ii), (b) [ Ph ]₃PAu]₃O⁺BF₄ ^-And [ Ph₃PAu]₂N⁺Concentration of R dimer and (c) [ Ph ]₃PAu]₃O⁺BF₄ ^-And [ Ph₃PAu]₃N⁺A plot of the results for the concentration of R trimer;

FIG. 5 is a graph of 100. mu. mol/LDA and 100. mu. mol/L [ Ph ] with various additional substrates added₃PAu]₃O⁺BF₄ ^-After derivatization, [ Ph ]₃PAu]⁺、[Ph₃PAu]₂N⁺R and [ Ph ]₃PAu]₃N⁺A calibration plot of the relative abundance of the R products;

in FIG. 6, (a) [ Ph ]₃PAu]₃O⁺BF₄ ^-Ultraviolet-visible absorption spectra of stroma in the presence of various neurotransmitters and (b-c) [ Ph₃PAu]⁺Transmission electron microscopy images of gold nanoparticles formed upon interaction with the neurotransmitter dopamine;

FIG. 7 shows dopamine and [ Ph ] at 100. mu. mol/L provided in example 1 of the present invention₃PAu]₃O⁺BF₄ ^-After derivatization of the matrix, forming a scanning electron microscope SEM image and an X-ray energy spectrum EDAX result of the gold nanoparticles;

FIG. 8 is a 50. mu. mol/L neurotransmitter mixture with 100. mu.M Ph provided in example 1 of the present invention₃PAu]₃O⁺BF₄ ^-MALDI-TOF-MS spectrogram after matrix derivatization;

FIG. 9 is a graph of (1) UV-visible absorption spectra of gold nanoparticles obtained at 527nm at various times as provided in example 1 of the present invention; (2) calibration plots between time and product abundance for different substrates;

FIG. 10 shows 100. mu. mol/L [ Ph ] provided in example 2 of the present invention₃PAu]₃O⁺BF₄ ^-MALDI-TOF-MS spectra in different regions of mouse brain extracts;

FIG. 11 is a calibration graph prepared by plotting the relative abundance and different neurotransmitters at different regions of the mouse brain, as provided in example 2 of the present invention.

Detailed Description

The present invention will be described in detail with reference to examples, but the present invention is not limited to these examples.

Unless otherwise specified, the raw materials used in the examples were purchased commercially and used without special treatment; in the examples, the equipment used was factory recommended parameters.

In the examples, energy dispersive x-ray spectroscopy (EDAX) elemental analysis and scanning electron microscopy topography (SEM) analysis used an in situ ultra high resolution field emission scanning electron microscope and energy spectrometer, model JSM-7800 (Prime).

The ultraviolet-visible absorption adopts a Thermo Scientific Evolution 201 type ultraviolet-visible spectrophotometer absorption spectrometer.

The matrix-assisted laser desorption ionization mass spectrometer is of the type MALDI-TOF/TOF 5800.

Example 1:

this example is the detection of mouse neurotransmitters, including Dopamine (DA), norepinephrine (NAd), 5-hydroxytryptamine (5-HT), gamma-aminobutyric acid (GABA), histamine (H).

Preparation of 1mmol/L [ Ph ]₃PAu]₃O⁺BF₄ ^-The acetonitrile solution of (1);

preparing 3nmol/L acetonitrile solutions of dopamine, norepinephrine, 5-hydroxytryptamine, gamma-aminobutyric acid and histamine respectively;

at 1mmol/L [ Ph ]₃PAu]₃O⁺BF₄ ^-Adding 0.2 mu L of each neurotransmitter solution into 1 mu L of the acetonitrile solution to form a mixed solution;

the mixed solution was stirred with a vortex shaker for 20 minutes. After 20 minutes, the mixed solution was transferred out of the vortex shaker and the reaction was continued for 3 hours. When a wine red colloidal suspension was observed, the formation of gold nanoparticles was confirmed;

the obtained sample was washed by centrifugation at 12000rpm for 5 minutes. After centrifugation, the supernatant was collected and dropped onto MALDI plates for analysis after drying.

The results confirmed by MALDI-TOF-MS at [ Ph₃PAu]₃O⁺BF₄ ^-The formation of Au NCLs with various neurotransmitters added. According to the results obtained, [ Ph ]₃PAu]₃O⁺BF₄ ^-And DA, NAd and 5-HT were derivatized to form a mixture of Au dimers and trimers, with dimers being more abundant than trimers and no monomer being found. And [ Ph₃PAu]₃O⁺BF₄ ^-And H derivatization produces monomers, dimers, and trimers. The abundance of monomer is higher than that of dimer and trimer.

In addition, catechols are neurotransmittersThe substances can also be used as reducing agents to convert Au NCLs into wine red Au NPs. The catechol neurotransmitters readily autooxidize in air to form intermediate semiquinones and/or quinones that release one or two electrons. As shown in fig. 2, the released electrons can reduce the Au ions to neutral Au NPs. And the formation of Au NPs was observed only in catecholamine neurotransmitters such as DA, NAd and 5-HT, etc., while the formation of Au NPs was not found in non-catecholamine neurotransmitters such as H and GABA. As shown in FIG. 6a, the formation of Au NPs was confirmed by UV-visible absorption bands at 527nm and 300 nm. The absorption band at 527nm shows the presence of metallic Au NPs, while the other absorption band at 300nm shows [ Ph₃PAu]₃O⁺BF₄ ^-The matrix is a phenylphosphine proton peak on the Au surface. The absorbance peak at 527nm was not detected for the non-catecholamines, whereas the non-catecholamines (GABA) retained only the phenylphosphinic peak.

As shown in fig. 6b and 6c, neurotransmitter is transmitted through [ Ph₃PAu]₃O⁺BF^4-Transmission Electron Microscopy (TEM) images of spherical gold nanoparticles formed after matrix derivatization. FIG. 7 is a graph of warp [ Ph ]₃PAu]₃O⁺BF^4-And (3) performing element analysis on Scanning Electron Microscope (SEM) images and energy spectrum x-ray spectra (EDAX) of the gold nanoparticles formed after derivatization of the matrix. SEM images show gold nanoparticles, EDAX evidencing the presence of gold atoms from my samples.

Some other triphenylphosphine may also be used to introduce [ Ph ] onto a particular substrate₃PAu]⁺Elements including phosphinofluoroborate, halides, trifluorides, perchlorates, and acetoacetates. To this end, we have selected [ Ph₃PAu]₃O⁺BF₄This is because the high affinity of the oxide function is believed to be a useful driving force for the formation of Au NCLs by the ammonium derivative. [ Ph₃PAu]₃O⁺BF₄The oxygen ion in (E) is also a template for the ammonium derivative. [ Ph₃PAu]₃O⁺BF₄The oxygen ion and Au ion in-play the same role in forming an ammonium derivative with the primary amine group of neurotransmitter. Au ion bonding by providing strong Au-N-Au bondTo increase the bonding strength of the ammonium derivative, while the oxygen ion provides the template for such derivatization. [ Ph₃PAu]₃O⁺BF₄Ph in (1)₃The P ligand inhibits the aggregation tendency of Au ions and provides a stable structure for Au clusters and nanoparticles. To investigate this hypothesis, oxygen ion-free chlorotriphenylphosphine gold ([ Ph)₃PAu]Cl^-) And trimethyl ammonium oxide tetrafluoroborate ([ CH ] containing no Au ion₃]₃O⁺BF₄ ^-) Is substituted by [ Ph₃PAu]₃O⁺BF₄ ^-. Unfortunately, no composition containing [ Ph ] has been found₃PAu]Cl and [ CH₃]₃O⁺BF₄ ^-Any product of neurotransmitter DA. This is due to [ Ph₃PAu]Cl^-Middle lack of oxygen ion template, [ CH₃]₃O⁺BF₄ ^-Due to the lack of Au ions. It has also been found that [ CH₃]₃O⁺BF₄ ^-Unstable at room temperature because [ CH ] is not observed in the mass spectrum₃]₃O⁺BF₄ ^-Characteristic peak of (2).

The simultaneous detection of various neurotransmitters such as DA, NAd, 5-HT, GABA and H was investigated by using MALDI-TOF-MS spectra, as shown in FIG. 8. By mixing 50. mu. mol/L of each neurotransmitter with 100. mu. mol/L of the same [ Ph ]₃PAu]₃O⁺BF₄ ^-The matrix solutions are mixed to prepare the sample for simultaneous detection. According to the results obtained, DA, NAd, 5-HT, GABA and H with [ Ph ] were observed in the same MALDI-TOF-MS spectrum₃PAu]₃O⁺BF₄ ^-The product of (1). Although [ Ph₃PAu]₃O⁺BF₄ ^-Au monomers, dimers, and trimers are formed for a variety of neurotransmitters, but Au dimers are generally obtained for all neurotransmitters. Therefore, we used only Au dimers for simultaneous detection of various neurotransmitters. The peaks of GABA-H, H-DA, DA-NAd and NAd-5-HT are well separated, so that the embodiment of the invention can simultaneously perform qualitative and quantitative detection on DA, NAd, 5-HT, GABA and HAnd (6) measuring.

The response time for Au NPs formation was further investigated by measuring uv-visible absorbance at 527nm wavelength, as shown in fig. 9 a. The ultraviolet-visible spectrum result shows that the absorbance is kept unchanged between 0.5min and 3min after the catecholamine neurotransmitter is added, and is gradually increased after 3 min. The results show that Au was formed 3min before DA addition⁰Layer, referred to as nucleation process. After 3min, Au⁰The nucleation layer serves as a seed layer for the propagation of Au NPs, which is called a growth process. Within the first 0.5min after neurotransmitter addition, [ Ph₃PAu]The concentration drop, followed by dilution, is probably due to [ Ph ]₃PAu]The concentration is reduced. The results also show that NAd with three hydroxyl groups has a faster response than DA with two hydroxyl groups, whereas 5-hydroxytryptamine reacts very slowly.

The results show that the reaction time is proportional to the number of hydroxyl groups in the neurotransmitter that act as electron donors. Catecholamine neurotransmitter with more hydroxyl groups releases more electrons in the oxidation process, and the electron group of reduction of Au ions into Au NPs is increased. Further, [ Ph ]₃PAu]₃O⁺BF₄ ^-The response times of (a) were also compared to previously reported reaction times for pyridinium salts such as 2, 4, 5-triphenylpyridinium Tetrafluoroborate (TPP), 2, 4, 6-trimethylpyridinium Tetrafluoroborate (TMP), and 2, 6-diphenylpyridinium tetrafluoroborate (DPP) as shown in FIG. 9 b. The results show that [ Ph₃PAu]₃O⁺BF₄ ^-Is 10 times faster than pyridine.

The detection Limits (LOD) of DA, NAd, 5-HT, GABA and H in the range of several nanospheres were determined using the 3S/N formula, which is sufficiently sensitive to neurotransmitters at both cellular and tissue levels. Separating GABA-H, H-DA, DA-NAd and NAd-5-HT from the obtained MALDI-TOF-MS peak, wherein GABA-H8.0311 Da, H-DA 41.9953Da, DA-NAD 15.975Da and NAD-5-HT 7.0232 Da; for the selective simultaneous detection of DA, NAd, 5-HT, GABA and H, the response time during formation of Au NPs was 30 seconds, and the neurotransmitter was rapidly detected, as shown in FIG. 10. The results show that our [ Ph ]₃PAu]₃O⁺BF₄ ^-Can successfully treatThe method is used for detecting the neurotransmitter in the animal brain tissue extract by MALDI-TOF-MS. The detection results are shown in table 1:

table 1 test results of example 1

Kind of neurotransmitter	Dopamine	Norepinephrine	5-hydroxytryptamine	Gamma-aminobutyric acid	Histamine
						Concentration (nmol/L)	3.0048	3.0033	3.0024	3.0022	2.9991

Example 2:

this example is similar to example 1, except that MALDI-TOF-MS detection of neurotransmitters in mouse brain sample extracts was performed.

One year old male Kunming mice (20-22g) were housed in individually ventilated cages and kept in an environment with free access to standard food and water, at 22 ℃ and 50% humidity. All experiments were performed according to the guidelines of the Chinese society for laboratory animal science to ensure animal welfare.

Mouse brain extracts were prepared as follows:

after the mice are anesthetized with 100mg/kg chloral hydrate solution, the animals are killed by decapitation, the heads are taken out immediately, and the mice are heated to 80 ℃ within 16 seconds by microwave radiation;

the brain was quickly removed from the cranium and three areas (striatum, hippocampus and prefrontal cortex) were dissected. Tissues from each area were collected from the brains of five different animal mice, pooled together, homogenized with an automated mini bead stirrer (from Biospec products), and divided into three separately processed samples;

striatum, hippocampus and frontal lobe cortex tissues (90 mg each) were homogenized separately with ice cold acetonitrile 300 μ L containing 0.3% formic acid. Homogenization was performed five times for a duration of 30 seconds (10 seconds for each interval);

the homogenate was centrifuged at 12000rpm for 30 minutes at 4 ℃. The supernatant was collected and filtered through a 0.20 μm filter to obtain a supernatant, i.e., a sample solution, for use.

Take 1mmol/L [ Ph ]₃PAu]₃O⁺BF₄ ^-To 100. mu.L of the acetonitrile solution of (3), 100. mu.L of the sample solution was added to form a mixed solution.

The mixed solution was examined in the same manner as in example 1.

The MALDI-TOF-MS result is shown in FIG. 10. Extracts of the striatum, hippocampus and prefrontal cortex were used for neurotransmitter analysis. In [ Ph₃PAu]₃O⁺BF₄ ^-MALDI-TOF-MS samples were prepared by adding different brain extracts to the solution. As shown in FIG. 11, neurotransmitters DA, NAd, 5-HT, GABA and TY were successfully detected in all brain regions, while H was not detected anywhere. It was also shown that hippocampal regions have higher abundance of DA, 5-HT and GABA, while prefrontal cortex and striatum have higher abundance of DA, 5-HT and GABA. In addition, higher abundance of TY was also found in all samples. Indicating that the concentration of neurotransmitters in the brain is regionally dependent.

In summary, organometallic [ Ph ] was detected by MALDI-TOF-MS₃PAu]₃O⁺BF₄ ^-The neurotransmitter in the brain tissue extract can be detected. [ Ph₃PAu]₃O⁺BF₄ ^-Provides a method for rapidly and simultaneously detecting neurotransmitters DA, NAd, 5-HT, GABA, H, TY and the like on a tissue level. [ Ph₃PAu]₃O⁺BF₄ ^-Catecholamine neurotransmitters DA, NAd and 5-HT are also visually distinguished from non-catecholamine neurotransmitters (GABA and H) by a color change. In addition, the concentrations of neurotransmitters in brain regions such as striatum, prefrontal cortex and hippocampus were also different, confirming that the concentration of neurotransmitters in brain is regional.

The above description is only for the purpose of illustrating the present invention and is not intended to limit the present invention in any way, and the present invention is not limited to the above description, but rather should be construed as being limited to the scope of the present invention.

Claims (10)

1. A laser desorption ionization mass spectrometry matrix comprising gold tris (triphenylphosphine) tetrafluoroborate.

2. A mass spectrometer, wherein the mass spectrometer is a matrix-assisted laser desorption ionization mass spectrometer; the matrix comprises gold tris (triphenylphosphine) tetrafluoroborate.

3. A detection method is characterized in that a substance to be detected in the detection method is selected from polypeptides containing primary amine groups;

the detection method uses matrix-assisted laser desorption ionization mass spectrometry; the matrix comprises gold tris (triphenylphosphine) tetrafluoroborate.

4. The detection method according to claim 3, wherein the molecular weight of the primary amine group-containing polypeptide is less than 500.

5. The detection method according to claim 3, wherein the substance to be detected is at least one selected from the group consisting of dopamine, norepinephrine, 5-hydroxytryptamine, γ -aminobutyric acid, histamine, and tyramine.

6. Detection method according to claim 3, characterized in that it comprises at least the following steps:

dissolving a tris (triphenylphosphine) gold tetrafluoroborate matrix and a substance to be detected in an organic solvent respectively to form a matrix solution and a solution to be detected;

and mixing the matrix solution and the solution to be detected to form a mixed solution, spotting the mixed solution on a matrix-assisted laser desorption ionization target plate, and analyzing by using the matrix-assisted laser desorption ionization mass spectrometry after drying.

7. The detection method according to claim 6, wherein the organic solvent is at least one selected from methanol, acetone, isopropanol and acetonitrile.

8. The detection method according to claim 6, wherein the molar ratio of the tris (triphenylphosphine) gold tetrafluoroborate to the sample to be detected in the mixed solution is greater than 10: 1.

9. detection method according to claim 3, characterized in that it comprises at least the following steps:

dissolving a tris (triphenylphosphine) gold tetrafluoroborate matrix and a substance to be detected in an organic solvent respectively to form a matrix solution and a solution to be detected;

mixing the matrix solution and the sample solution, and centrifuging to obtain a supernatant;

and (3) spotting the supernatant on a matrix-assisted laser desorption ionization target plate, and performing mass spectrometry after drying.

10. Use of the mass spectrometer of claim 2 for neurotransmitter detection.

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