CN112924534B - Preparation method of nano bismuth/graphene composite material and application of nano bismuth/graphene composite material in MALDI-MS - Google Patents

Preparation method of nano bismuth/graphene composite material and application of nano bismuth/graphene composite material in MALDI-MS Download PDF

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CN112924534B
CN112924534B CN202110142044.8A CN202110142044A CN112924534B CN 112924534 B CN112924534 B CN 112924534B CN 202110142044 A CN202110142044 A CN 202110142044A CN 112924534 B CN112924534 B CN 112924534B
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bismuth
composite material
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graphene composite
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CN112924534A (en
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唐雪妹
万凯
陈志廷
黄健祥
高毓文
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Guangdong Agricultural Monitoring Technology Co ltd
Institute Of Agricultural Quality Standards And Monitoring Technology Guangdong Academy Of Agricultural Sciences
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Institute Of Agricultural Quality Standards And Monitoring Technology Guangdong Academy Of Agricultural Sciences
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Abstract

The invention relates to the technical field of analysis, in particular to a preparation method of a nano bismuth/graphene composite material and application of the nano bismuth/graphene composite material in MALDI-MS, which comprises the following steps: s01, preparing GO, and preparing a GO water solution with the concentration of 0.5-1.5 mg/mL; s02, adding bismuth salt in the step S01, stirring for 20-40 min, and adjusting the pH value of the solution to 9-10; s03, placing the solution treated in the step S02 in a reaction kettle to react for 8-12 hours; and S04, washing the solid obtained by the reaction in the step S03 with ethanol and water for several times, and then drying in vacuum to obtain the bismuth/graphene composite material. The invention constructs a MALDI-MS negative ion mode analysis method based on bismuth-based nano materials, and can influence the electron distribution in the system by constructing the composite material, thereby increasing the light absorption of the composite material, accelerating the separation of electron-hole pairs and improving the dissociation efficiency of the object to be detected.

Description

Preparation method of nano bismuth/graphene composite material and application of nano bismuth/graphene composite material in MALDI-MS
Technical Field
The invention relates to the technical field of analysis, in particular to a preparation method of a nano bismuth/graphene composite material and application of the nano bismuth/graphene composite material in MALDI-MS.
Background
Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry has the advantages of high throughput, high sensitivity, high salt tolerance, high analysis speed, small sample consumption, simple preparation process and the like, and has become one of the technical means widely applied in the field of mass spectrometry. MALDI mass spectrometry has become well established in the study of macromolecules such as polypeptides, proteins, nucleic acids, etc., however it still presents significant challenges in the detection of small molecule compounds. Severe background interference of the matrix, non-uniform matrix coverage and crystallization cause signal instability, poor dot-dot reproducibility, low ionization efficiency in the negative ion mode, and the like, which limits the wide application of the compound with low molecular weight. The small molecular substances cover a wide range of amino acids, polypeptides, metabolites, hormones and the like related to a life system, drugs related to the medical field, pollutants related to the environmental field and the like, and the rapid and effective analysis of the small molecular substances is significant. With the increasing attention paid to the research of matrix-assisted laser ionization mass spectrometry (MALDI) in the field of small molecule compounds, how to eliminate matrix interference, improve detection sensitivity and expand application range becomes a problem with great practical significance.
Over the last decade, scientific researchers have made extensive efforts to develop different matrices and methods for MALDI studies of low molecular weight compounds. The method of using a plurality of matrixes in combination, synthesizing a high molecular weight matrix, developing a novel ionic compound matrix and the like improves the background interference problem of the matrixes to a certain extent, but the problems of uneven crystallization of the matrixes and the samples, poor reproducibility and the like are not completely solved. Most of the existing researches are focused on developing various inorganic materials for positive ion mode mass spectrometry identification according to the thermal drive dissociation mechanism of nano materials, however, various alkali metal ion adducts can be generated under the positive ion mode, the spectrogram is complex, and the identification difficulty is increased.
Disclosure of Invention
The invention aims to overcome the problems in the prior art and provides a preparation method of a nano bismuth/graphene composite material and MALDI-MS analysis of the nano bismuth/graphene composite material in a negative ion mode.
The purpose of the invention is realized by the following technical scheme:
a preparation method of a nano bismuth/graphene composite material comprises the following steps:
s01, preparing GO, and preparing a GO water solution with the concentration of 0.5-1.5 mg/mL;
s02, adding bismuth salt into the solution in the step S01, wherein the concentration of the bismuth salt is 30mM, stirring for 20-40 min, and adjusting the pH value of the solution to 9-10;
s03, placing the solution treated in the step S02 in a reaction kettle to react for 8-12 hours at the temperature of 180 ℃;
and S04, washing the solid obtained by the reaction in the step S03 with ethanol and water for several times, and then drying in vacuum to obtain the bismuth/graphene composite material.
The nano bismuth/graphene composite material (Bi)2O3@ GO) is prepared.
Nanomaterials are of great interest due to their own unique optical properties and thermal and electrical conductivity properties. Graphite materials, porous silicon, metal oxides, metal particles, metal organic frameworks and other materials have been widely used as mass spectrometry ionization matrices to realize detection and analysis of multi-component small molecule mixtures. Compared with the traditional matrix, the substance has unique advantages, such as avoiding the problem of uneven crystallization of the matrix, reducing the ion inhibition effect of the matrix on a sample, and the like. The application of the nano material enhances the signal intensity of the sample to a certain extent, widens the application of MALDI mass spectrometry in the field of small molecular compounds, and improves the detection sensitivity.
The application of a bismuth-based nano material in MALDI-MS is provided, wherein the bismuth-based nano material comprises bismuth oxide nano or the bismuth/graphene composite material.
Preferably, the application of the bismuth-based nano material in MALDI-MS comprises the following steps:
s11, dispersing the bismuth-based nano material in an organic solvent, and performing ultrasonic treatment to form a suspension of 1-10 mg/mL;
s12, dropwise adding the suspension obtained in the step S11 on a MALDI metal target and air-drying;
s13, dropwise adding the solution to be detected on the surface of the nano material obtained in the step S12, and performing MALDI-MS detection after air drying.
Compared with the prior art, the invention has the following technical effects:
the bismuth-based nano material is used in MALDI-MS detection, has the advantages of narrow forbidden band width, high stability, low cost, environmental friendliness and the like, can absorb the energy excited by ultraviolet laser from matrix-assisted laser desorption ionization time-of-flight mass spectrometry, and promotes desorption ionization of an object to be detected. The material does not generate mass spectrum peak, can effectively overcome the matrix interference problem of the traditional organic matrix to the detection of the micromolecular substance, and can be suitable for carrying out mass spectrum analysis on the micromolecule with the molecular weight less than 1000. In addition, the invention constructs a MALDI-MS negative ion mode analysis method based on the bismuth-based nano material, and can influence the electron distribution in the system by constructing the composite material, thereby increasing the light absorption of the composite material, accelerating the separation of electron-hole pairs and improving the dissociation efficiency of the object to be detected.
Drawings
FIG. 1 is an SEM image of a bismuth oxide nanomaterial;
fig. 2 is an SEM image of the bismuth-graphene composite material;
FIG. 3 shows the results of measuring 10mM glucose signal using bismuth-based nanomaterial as a substrate ((A) Bi)2O3(B)Bi2O3@GO)；
FIG. 4 shows a Bi-based nanomaterial Bi2O3Detecting the analysis result of 10mM mixed amino acid solution by using @ GO as a matrix;
FIG. 5 shows Bi-based nanomaterial Bi2O3Detecting analysis results of five 200nM mixed fatty acid solutions by using @ GO as a matrix;
FIG. 6 shows Bi-based nanomaterial Bi2O3@ GO is used as a matrix for detecting the analysis result of the small molecule metabolite.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the technical solutions of the present invention will be described in detail below with reference to specific examples and comparative examples. It should be apparent that the described embodiments are only some embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the examples given herein without any inventive step, are within the scope of the present invention.
Unless otherwise specified, the devices used in this example are all conventional experimental devices, the materials and reagents used are commercially available, and the experimental methods without specific descriptions are also conventional experimental methods.
Example 1
A preparation method of a nano bismuth/graphene composite material comprises the following steps:
s01, preparing GO, and preparing a GO water solution with the concentration of 0.5-1.5 mg/mL;
s02, adding bismuth salt in the step S01, wherein the concentration of the bismuth salt is 30mM, stirring for 20-40 min, and adjusting the pH value of the solution to 9-10;
s03, placing the solution treated in the step S02 in a reaction kettle to react for 8-12 hours at the temperature of 180 ℃;
and S04, washing the solid obtained by the reaction in the step S03 with ethanol and water for several times, and then drying in vacuum to obtain the bismuth/graphene composite material.
The bismuth/graphene composite material is prepared by the preparation method of the nano bismuth/graphene composite material.
Preparing a bismuth oxide nano material: dispersing bismuth-containing salt in an organic solvent, and carrying out high-temperature high-pressure solvothermal reaction to obtain the required bismuth nano material. In the reaction process, the bismuth-containing salt solution can be a bismuth nitrate solution, the dispersing agent can be an ethylene glycol solution, and the specific process can be as follows: adding a certain amount of bismuth nitrate (Bi (NO)3)35H2O) was dispersed in 80ml of ethylene glycol and stirred for 1 hour, placed in a tetrafluoroethylene reactor, reacted at 60-200 ℃ for a certain time, the precipitate was collected, washed several times with pure ethanol and water and dried in a vacuum drying oven. The obtained nano material is ground and dispersed in isopropanol solution, and can be used as a matrix for MALDI-TOF MS detection. The particle size of the synthesized bismuth-based nano material is 50-200 nm.
The bismuth-based nano material (shown in an SEM figure in figure 1 and figure 2) is used as a matrix for matrix-assisted laser desorption ionization time-of-flight mass spectrometry detection, and the specific experimental process is as follows:
1. preparing 1.0-10mg/mL of organic suspension solution serving as a matrix solution from the obtained bismuth/graphene composite material;
2. performing ultrasonic treatment on the system at normal temperature for 1-2min to form a uniform dispersed suspension system;
3. preparing 10mM glucose sample solution as the sample solution to be tested
4. And dripping the nano suspension onto a MALDI-MS metal target plate, and drying the solution to form a uniform matrix layer.
5. And (3) dropwise adding the sample solution to the surface of the dried nano material in the step 3.
6. The dried spots were subjected to MALDI-TOF MS detection. All MALDI mass analyses were performed on an shimadzu imaging mass spectrometry microscope (ismscope TRIO) mass spectrometer. In the detection process, a mass spectrogram is obtained by using a laser under a 355nm negative ion mode, and the laser frequency is 1000 Hz. Each sample was randomly measured at 200 different spots, and each spot was randomly laser spotted 50 times.
As can be seen from FIG. 3, glucose can be detected well by MALDI-TOF MS, where glucose (glucose): M/z 179.06[ M-H ]. The results show that the matrix provided by the application can effectively detect the small molecular substances with the molecular weight less than 1000 Da.
Example 2
The bismuth oxide nanomaterial and the bismuth/graphene composite material were the same as in example 1.
The bismuth-based nano material is used as a matrix for matrix-assisted laser desorption ionization time-of-flight mass spectrometry detection, and the specific experimental process is as follows:
1. preparing 2.0mg/mL organic suspension solution of bismuth-based nano material as matrix solution;
2. performing ultrasonic treatment on the system at normal temperature for 1-2min to form a uniform dispersed suspension system;
3. five kinds of 10mM mixed amino acid solutions are prepared as sample solutions to be detected, the concentration is 10mM, and the specific amino acids are Asp, Glu, His, Arg and Try.
4. And dripping the nano suspension onto a MALDI MS metal target plate, and drying the solution to form a uniform matrix layer.
5. And (3) dropwise adding the sample solution to the surface of the dried nano material in the step 3.
6. The dried spots were subjected to MALDI-TOF MS detection. All MALDI mass analyses were performed on an shimadzu imaging mass spectrometry microscope (ismscope TRIO) mass spectrometer. In the detection process, a mass spectrogram is obtained by using a laser under a 355nm negative ion mode, and the laser frequency is 1000 Hz. Each sample was randomly measured at 200 different spots, and each spot was randomly laser spotted 50 times.
The results are shown in FIG. 4. As can be seen from fig. 4, when MALDI-TOF MS detection was performed using the matrices provided herein, each small amino acid, aspartic acid (Asp), M/z 132.03[ M-H ] -, of the mixed amino acids was detected; glutamic acid (Glu): M/z 146.05M-H ] -; histidine (His) M/z 154.06[ M-H ] -; arginine (Arg) M/z 173.10[ M-H ] -; tryptophan (Try): M/z 203.08[ M-H ] -. Therefore, the matrix provided by the application can effectively detect the small molecular substances with the molecular weight less than 1000 Da.
Example 3
The bismuth oxide nanomaterial and the bismuth/graphene composite material were the same as in example 1.
The bismuth-based nano material is used as a matrix for matrix-assisted laser desorption ionization time-of-flight mass spectrometry detection, and the specific experimental process is as follows:
1. preparing 2.0mg/mL organic suspension solution of bismuth-based nano material as matrix solution;
2. performing ultrasonic treatment on the system at normal temperature for 1-2min to form a uniform dispersed suspension system;
3. preparing 5 kinds of 200mM mixed fatty acid solutions as sample solutions to be tested, wherein the specific fatty acids are C12:0, C14:0, C16:0, C18:0 and C20:0
4. And dripping the nano suspension onto a MALDI MS metal target plate, and drying the solution to form a uniform matrix layer.
5. And (3) dropwise adding the sample solution to the surface of the dried nano material in the step 3.
6. The dried spots were subjected to MALDI-TOF MS detection. All MALDI mass analyses were performed on an shimadzu imaging mass spectrometry microscope (ismscope TRIO) mass spectrometer. In the detection process, a mass spectrogram is obtained by using a laser under a 355nm negative ion mode, and the laser frequency is 1000 Hz. Each sample was randomly measured at 200 different spots, and each spot was randomly laser spotted 50 times.
The results are shown in fig. 5, which shows that when MALDI-TOF MS detection is performed using the matrix provided herein, each small fatty acid in the mixed lipid can be detected, C12:0: M/z 199.17[ M-H ] -; c14:0: M/z 227.20[ M-H ] -; c16:0: M/z 255.23[ M-H ] -; c18:0: M/z 283.26[ M-H ] -; c20:0: M/z 311.29[ M-H ] -. Therefore, the matrix provided by the application can effectively detect the small molecular substances with the molecular weight less than 1000 Da.
Example 4
The bismuth oxide nanomaterial and the bismuth/graphene composite material were the same as in example 1.
The bismuth-based nano material is used as a matrix for matrix-assisted laser desorption ionization time-of-flight mass spectrometry detection, and the specific experimental process is as follows:
1. preparing 2.0mg/mL organic suspension solution of bismuth-based nano material as matrix solution;
2. performing ultrasonic treatment on the system at normal temperature for 1-2min to form a uniform dispersed suspension system;
3. preparing a mixed sample solution containing Ascorbic Acid (Ascorbic Acid), bisphenol A (BPA), exogenous drug Sulfaquinoxaline (Sulfaquinoxaline) and plant hormone Gibberellin (GA) with a concentration of 1.0mg/mL, and making into a sample solution
4. And dripping the nano suspension onto a MALDI MS metal target plate, and drying the solution to form a uniform matrix layer.
5. And (3) dropwise adding the sample solution to the surface of the dried nano material in the step 3.
6. The dried spots were subjected to MALDI-TOF MS detection. All MALDI mass analyses were performed on an shimadzu imaging mass spectrometry microscope (ismscope TRIO) mass spectrometer. In the detection process, a mass spectrogram is obtained by using a laser under a 355nm negative ion mode, and the laser frequency is 1000 Hz. 200 different spots were randomly measured for each sample, and each spot was randomly laser spotted 50 times.
As can be seen from fig. 6, when MALDI-TOF MS detection is performed using the matrix provided herein, wherein small molecule species can be well detected by MALDI-TOF MS, Ascorbic Acid (Ascorbic Acid): m/z 175.03[ M-H ] -; bisphenol A (BPA) as environmental pollutant, M/z is 227.11M-H-, M/z is 211.08M-OH-; exogenous drug Sulfaquinoxaline (Sulfaquinoxaline), M/z 299.06[ M-H ] -; plant hormone Gibberellin (GA): M/z is 345.14[ M-H ] -. The above results show that the matrix provided by the application can effectively detect small molecular substances with the molecular weight of less than 1000 Da.
Finally, it should be noted that the above embodiments are only used for illustrating the technical solutions of the present invention, and not for limiting the protection scope of the present invention, although the present invention is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions can be made on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.

Claims (1)

1. The application of the bismuth-based nano material in MALDI-MS is characterized in that the preparation method of the bismuth-based nano material comprises the following steps:
s01, preparing GO, and preparing a GO water solution with the concentration of 0.5-1.5 mg/mL;
s02, adding bismuth salt into the solution in the step S01, wherein the concentration of the bismuth salt is 30mM, stirring for 20-40 min, and adjusting the pH value of the solution to 9-10;
s03, placing the solution treated in the step S02 in a reaction kettle to react for 8-12 hours at the temperature of 180 ℃;
s04, washing the solid obtained by the reaction in the step S03 with ethanol and water for several times, and then drying in vacuum to obtain the bismuth graphene composite material;
the application method of the bismuth-based nano material in MALDI-MS comprises the following steps:
dispersing the bismuth-based nano material in an organic solvent, and performing ultrasonic treatment to form a suspension of 1-10 mg/mL;
s12, dropwise adding the suspension obtained in the step S11 on a MALDI metal target and air-drying;
s13, dropwise adding the solution to be detected on the surface of the nano material obtained in the step S12, air-drying, and then carrying out MALDI-MS detection.
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