CN111413394A - SiO2Application of core/Au shell nano material as matrix in MA L DI-TOF MS detection - Google Patents
SiO2Application of core/Au shell nano material as matrix in MA L DI-TOF MS detection Download PDFInfo
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Abstract
The invention provides SiO2The analysis method of the nano material as the MA L DI-TOF MS matrix is suitable for carrying out mass spectrum analysis on small molecules with the molecular weight less than 1000, greatly simplifies the detection difficulty of the small molecule sample, improves the detection sensitivity of the MA L DI-TOF MS of the small molecule sample, and simultaneously can realize the elimination of mass spectrum peak interference generated by the existing matrix and the enhancement processing of mass spectrum signals of the small molecule sample.
Description
Technical Field
The invention belongs to the technical field of mass spectrometry detection, and particularly relates to SiO2The application of the core/Au shell nano material as a matrix in MA L DI-TOF MS detection.
Background
Since metabolic alterations can directly participate in the transformation process or support the biological process of tumor growth, small molecule metabolites can become a unique source of cancer-specific information, and thus, the application of metabolomics in clinical tumor research has become a research hotspot in recent years. During this period, mass spectrometry also plays an increasingly important role.
Although the MA L DI-TOF MS is various, the organic small molecular matrix is difficult to be used for analyzing compounds with small molecular weight (less than 10000 Da), and the main reason is that the organic small molecular matrix is cracked and associated between molecules, so that a serious matrix background interference phenomenon is generated.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides SiO2The application of the core/Au shell nano material as a matrix in MA L DI-TOF MS detection is realized, and the analysis method of the nano material as the MA L DI-TOF MS matrix is suitable for carrying out mass spectrum analysis on small molecules with the molecular weight less than 1000, so that the detection difficulty of the small molecule sample is greatly simplified, the detection sensitivity of the MA L DI-TOF MS of the small molecule sample is improved, and simultaneously, the elimination of mass spectrum peak interference generated by the existing matrix and the enhancement processing of mass spectrum signals of the small molecule sample can be realized.
In order to achieve the purpose, the technical scheme adopted by the invention for solving the technical problems is as follows:
SiO (silicon dioxide)2The application of the core/Au shell nano material as a matrix in MA L DI-TOF MS detection.
Further, the molecular weight of the substance to be detected in the MA L DI-TOF MS detection is less than 1000.
Further, the substance to be detected in the MA L DI-TOF MS detection is a body fluid metabolite, and the body fluid metabolite can be serum, plasma, urine, sweat, semen, hydrocephalus, joint fluid and the like.
Further, the MA L DI-TOF MS detection is used for mass spectrum imaging of the substance to be detected.
Further, willDripping a body fluid sample containing metabolites onto a metal target sheet of MA L DI-TOF MS, and dripping SiO after the sample is dried by air2core/Au shell nanomaterials, after continued air drying, were subjected to MA L DI-TOF MS detection.
Further, SiO2The concentration of the core/Au shell nano material is 0.8-1.5mg/ml, SiO2The volume ratio of the core/Au shell nano material to the sample to be detected is 1-4: 1.
Further, SiO2The concentration of the core/Au shell nano material is 1mg/ml, SiO2The volume ratio of the core/Au shell nano material to the sample to be detected is 2: 1.
Further, SiO2core/Au shell nanomaterials (also known as Gold Nanoshell (GNS)) were prepared by the following method:
(1) preparation of gold nanoparticles
Chlorauric acid hydrate (HAuCl4 & 3H) was stirred2O) is added into the THPC solution, and then the THPC solution is placed at 4 ℃ for aging for 12-16h to prepare gold nanoparticles;
(2) preparation of functionalized silica core
Mixing the silicon dioxide nanospheres with ethanol, and uniformly dispersing by ultrasonic;
mixing the ultrasonic dispersion liquid with 3-aminopropyltriethoxysilane, performing heat treatment at 85-95 ℃ for 2-4h, cooling to room temperature, centrifuging, washing, and resuspending to obtain the final product;
(3) modification of silicon core by gold nano-particles
Mixing gold nanoparticles, functional silicon core and water according to a volume ratio of 1-2:1-2:2-5, standing at room temperature for 8-12min, standing at 4 ℃ for 24-28h, cleaning, and resuspending to obtain the final product;
(4) growth of gold nanoshell
Mixing carbonate solution and chlorauric acid hydrate (HAuCl4 & 3H)2O) to obtain a colorless solution, then placing the colorless solution at 4 ℃ for aging for 24-28h, then adding the aged solution into the product obtained in the step (3) while stirring, adding a reducing agent after reacting for 8-15min, and continuing stirring until the reduction is complete to obtain the gold nano shell layer.
Further, the THPC solution in step (1) is prepared by the following method: 1M of sodium hydroxide, THPC and water are mixed according to the volume ratio of 0.4-0.6: 10-15: 45-50, and mixing; wherein the volume ratio of 1M sodium hydroxide, THPC and water is preferably 0.5: 12: 47.5.
further, the specific steps in the step (2) are as follows: mixing 2.5 wt% of silicon dioxide nanospheres with ethanol, uniformly dispersing by ultrasonic, adding 3-aminopropyl triethoxysilane, carrying out heat treatment at 90 ℃ for 3h, cooling to room temperature, centrifuging, washing with ethanol, and finally carrying out heavy suspension with ethanol to obtain the silica nanospheres; wherein, the volume ratio of the 2.5 wt% silicon dioxide nanosphere, the ethanol and the 3-aminopropyl triethoxysilane is 0.3-0.6:8-12:0.1-0.3, preferably 0.5:10: 0.15.
Further, in the step (3), the volume ratio of the gold nanoparticles to the functionalized silicon core to the water is 1:1: 3.
Further, the volume ratio of the aged solution in the step (4), the product obtained in the step (3) and the reducing agent is 6-10:18-25:45-55, preferably 8:20: 50.
Further, in the step (4), the carbonate is potassium carbonate, and the reducing agent is formaldehyde.
The SiO provided by the invention2The application of the core/Au shell nano material as a matrix in MA L DI-TOF MS detection has the following beneficial effects:
according to the invention, a gold shell grows on a silicon core, and a continuous nano shell is grown in situ by absorbing gold nanoparticles synthesized in advance on silicon beads, so that a core-shell nanosphere, namely a gold nano shell layer is formed, the gold nano shell layer has a rough surface and a strong surface plasma effect, so that the gold nano shell layer also shows strong absorption in an ultraviolet region, and the unique performance is favorable for absorbing energy excited by ultraviolet laser from matrix-assisted laser desorption ionization flight time mass spectrum to a great extent, so that a small molecular metabolite signal smaller than 1000Da can be captured and analyzed, laser energy can be absorbed without generating detectable cluster ions, the defect that the existing matrix easily generates a serious matrix background interference phenomenon in a low molecular weight region, so that a small molecular sample cannot be effectively analyzed is overcome, the detection difficulty of the small molecular sample is greatly simplified, and the detection sensitivity of MA L DI-MS of the small molecular sample is improved.
Drawings
FIG. 1 is a graph showing the results of characterization of Gold Nanoshells (GNS).
FIG. 2 is SiO2Schematic diagram of core/Au shell nano material as matrix in MA L DI-TOF MS detection.
FIG. 3 is SiO2Typical L DI signal results after mixing of core/Au shell nanomaterials as matrices with serum samples.
FIG. 4 is SiO2And the core/Au shell nano material is used as a substrate to be mixed with a serum sample, and then a glucose signal result is detected.
FIG. 5 is SiO2And the core/Au shell nano material is used as a substrate to be mixed with a serum sample, and then the glucose and uric acid signal results are detected.
FIG. 6 shows the results of the measurement of ten mixed amino acids at a concentration of 200 nM.
FIG. 7 shows the results of the measurement of ten mixed amino acids at a concentration of 1000 nM.
FIG. 8 shows the results of detection at a concentration of 0.25. mu.M for four mixed amino acids.
FIG. 9 shows the results of detection at a concentration of 2.5. mu.M for four mixed amino acids.
FIG. 10 shows the results of detection at a concentration of 5. mu.M for four mixed amino acids.
FIG. 11 shows the results of detection at a concentration of 25. mu.M for four mixed amino acids.
Detailed Description
EXAMPLE 1 preparation of gold nanoshell layer
(1) Preparation of gold nanoparticles
Mixing 0.5ml of 1M sodium hydroxide, 12ml of THPC and 47.5ml of water to prepare a THPC solution;
2.06m L of 1 wt% chloroauric acid hydrate (HAuCl4 & 3H) were stirred2O) is rapidly added into the THPC solution, the covering layer can be observed to turn brown within 1 minute, which indicates the formation of gold nano colloid, and then the gold nano colloid is stored and aged at 4 ℃ for at least 12 hours for use;
(2) preparation of functionalized silica core
Mixing 0.5ml of 2.5 wt% silica nanosphere (120nm, Tianjin Dart technologies, Ltd.) with 10ml of ethanol, and ultrasonically dispersing for 2min to uniformly disperse; then adding 150ul 3-aminopropyl triethoxysilane (APTES), heat treating at 90 deg.C for 3h, cooling to room temperature, centrifuging at 5000r/min for 10min, washing in ethanol for 3 times, and finally resuspending with 2ml ethanol to obtain the final product;
(3) modification of silicon core by gold nano-particles
Mixing 2ml of gold nanoparticles, 2ml of functional silicon core and 6ml of water, standing at room temperature for 10min, transferring to 4 ℃, standing for at least 24h to obtain gold particle-silicon nanospheres, finally cleaning the gold particle-silicon nanospheres for 3 times, and carrying out heavy suspension by using 2ml of water to obtain the gold particle-silicon nanospheres;
(4) growth of gold nanoshell
25mg of potassium carbonate are dissolved in 100ml of deionized water, and 2ml of 1 wt.% HAuCl are added4·3H2Chloroauric acid O hydrate (HAuCl4 & 3H2O) mixing to obtain a colorless solution, then placing the colorless solution at 4 ℃ for aging for at least 24h, then adding 8ml of the aged solution into 20ul of the product obtained in the step (3) while stirring, reacting for 10min, then slowly supplementing 50 mu L formaldehyde (37%) serving as a reducing agent into the solution, continuing stirring for 24h, and at the moment, completely reducing to obtain a gold nano shell (namely SiO nano shell)2core/Au shell nanomaterial).
Scanning Electron Microscope (SEM), Transmission Electron Microscope (TEM) and ultraviolet-visible light (UV-vis) detection are carried out on the prepared gold nanoshell layer, and the detection result is shown in figure 1. Wherein, fig. 1a is a Scanning Electron Microscope (SEM) image of the gold nanoshell layer (GNS), fig. 1b is a Transmission Electron Microscope (TEM) image of the gold nanoshell layer (GNS), and fig. 1c is an ultraviolet-visible light absorption spectrum image of the gold nanoshell layer (GNS).
As can be seen from FIG. 1a, the gold nanoshell has a uniform particle size distribution, a size of about 120nm, and a relatively rough surface compared to a smooth surface of the bare silicon nanoparticle, which is caused by particle collapse induced by different gold nanoparticle seeds.
As can be seen from fig. 1b, the element distributions of O (sky blue), Si (red) and Au (yellow) were analyzed by TEM image and EDX, and it was clearly confirmed that the core-shell structure was formed, the core contained abundant Si and O elements, the shell contained abundant Au elements, and the shell was about 10nm thick.
As can be seen from fig. 1c, the plasmon peak of the gold nanoshell nanoparticles is about 680nm, the uv absorption spectrum is about 270nm, and the uv-vis absorption peak around 680nm also reveals the formation of a gold shell on the insulating core structure.
Example 2 matrix-assisted laser desorption ionization time-of-flight mass spectrometry
Detecting small molecular substances in serum
An experimental group is that 0.5u L serum sample is dripped on a polished steel target plate MTP384, after air drying, 1u L Gold Nano Shell (GNS) nano material is dripped, the concentration of the Gold Nano Shell (GNS) nano material is 1mg/ml, after complete drying, the sample is arranged on an Autoflex Max mass spectrometer (Bruker Daltonics company of Bremen, Germany) to carry out mass spectrum detection, in the detection process, a smartpeak-II laser is used for obtaining a spectrum under a 355nm reflection positive mode, each sample with the laser frequency of 1000 hz. randomly measures 20 different points, and each point is subjected to laser random dotting for 25 times, so that 500 satisfactory dotting can be obtained.
In the control group, the Gold Nanoshell (GNS) nanomaterial was replaced with α -cyano-4-hydroxyphenylacrylic acid (CHCA) matrix, and the other procedures were the same as those in the experimental group.
SiO2A schematic diagram of core/Au shell nanomaterial (GNS) as a matrix in MA L DI-TOF MS detection is shown in FIG. 2, wherein SiNW (silicon nanowire) in FIG. 2 is also a matrix.
SiO2The results of a typical L DI signal after mixing the core/Au shell nanomaterial as a matrix with a serum sample are shown in FIG. 3, where the curves in FIG. 3 represent CHCA × 10, GNS, and SiNW, respectively, from bottom to top.
As can be seen from FIG. 3, the serum sample mixed with GNS or SiNW produced significant signals at m/z of 100-900Da after passing through the mass spectrometer, and the results of the mass spectrometer were partly detailed in the following manner, glucose (G L U), m/z 181.07[ G L U + H ]]+Glucose (G L U) m/z 203.05[ G L U + Na ]]+Glucose (G L U) m/z 219.026[ G L U + K ]]+(ii) a Uric Acid (UA): 169.035[ UA + H ] m/z]+(ii) a Uric Acid (UA): m/z 191.017[ UA + Na]+(ii) a Uric Acid (UA): m/z 206.99[ UA + K]+(ii) a Creatinine (CRE): 114.06[ CRE + H ] m/z]+(ii) a Creatinine (CRE): 136.048[ CRE + Na ] m/z]+(ii) a Creatinine (CRE): m/z 152.02[ CRE + K ═]+(ii) a Partial results are shown in fig. 4 and 5.
In contrast, mixtures that directly mix conventional α -cyano-4-hydroxyphenylacrylic acid (CHCA) matrices with serum samples gave little if any signal, even a 10-fold expansion, which may be due to lipid interference.
Secondly, detecting signals in the mixture of ten amino acids
The detection was performed according to the detection method in the above experimental group, except that the serum sample was replaced with ten mixed amino acids at a concentration of 200nM, and the specific amino acids were Ser, Pro, Thr, L eu, Met, L ys, His, Phe, Try, and Val, and the detection results are shown in FIG. 6.
The amino acid concentration was changed to 1000nM, and the results are shown in FIG. 7.
As can be seen from FIGS. 6 and 7, when MA L DI-TOF MS detection was performed using the matrix provided herein, each small-molecule amino acid among the mixed amino acids was detected, and thus it was found that the matrix provided herein was effective for detecting small-molecule substances having a molecular weight of less than 1000 Da.
Thirdly, detecting signals in the mixture of four amino acids
The detection was performed according to the detection method in the above experimental group, except that the serum sample was changed to four mixed amino acids at a concentration of 0.25. mu.M, and the specific amino acids were Met, Phe, Try and Thr, and the detection results are shown in FIG. 8.
The amino acid concentration was changed to 2.5. mu.M, and the results are shown in FIG. 9.
The amino acid concentration was changed to 5. mu.M, and the results are shown in FIG. 10.
The amino acid concentration was changed to 25. mu.M, and the results are shown in FIG. 11.
From FIGS. 8-11, it can be seen that each of the small amino acids in the mixture of amino acids can be detected when MA L DI-TOF MS detection is performed using the matrix provided herein, and thus the matrix provided herein can effectively detect small substances having a molecular weight of less than 1000 Da.
Claims (10)
1. SiO (silicon dioxide)2The application of the core/Au shell nano material as a matrix in MA L DI-TOF MS detection.
2. Use according to claim 1, wherein the molecular weight of the substance to be detected in the MA L DI-TOF MS assay is less than 1000.
3. The use according to claim 1 or 2, wherein the substance to be tested in the MA L DI-TOF MS assay is a body fluid metabolite.
4. Use according to claim 1, wherein the MA L DI-TOF MS detection is used for mass spectrometric imaging of a substance to be tested.
5. Use according to claim 1, wherein the metabolite-containing body fluid sample is applied dropwise to a metal target of MA L DI-TOF MS, and after the sample has dried, SiO is added dropwise2core/Au shell nanomaterials, after continued air drying, were subjected to MA L DI-TOF MS detection.
6. Use according to claim 1 or 5, wherein the SiO is2The concentration of the core/Au shell nano material is 0.8-1.5mg/ml, SiO2The volume ratio of the core/Au shell nano material to the sample to be detected is 1-4: 1.
7. Use according to claim 6, wherein the SiO is2The concentration of the core/Au shell nano material is 1mg/ml, SiO2The volume ratio of the core/Au shell nano material to the sample to be detected is 2: 1.
8. Use according to claim 1, wherein the SiO is2The core/Au shell nano-material is prepared by the following methodThe method comprises the following steps:
(1) preparation of gold nanoparticles
Chlorauric acid hydrate (HAuCl4 & 3H) was stirred2O) is added into the THPC solution, and then the THPC solution is placed at 4 ℃ for aging for 12-16h to prepare gold nanoparticles;
(2) preparation of functionalized silica core
Mixing the silicon dioxide nanospheres with ethanol, and uniformly dispersing by ultrasonic;
mixing the ultrasonic dispersion liquid with 3-aminopropyltriethoxysilane, performing heat treatment at 85-95 ℃ for 2-4h, cooling to room temperature, centrifuging, washing, and resuspending to obtain the final product;
(3) modification of silicon core by gold nano-particles
Mixing gold nanoparticles, functional silicon core and water according to a volume ratio of 1-2:1-2:2-5, standing at room temperature for 8-12min, standing at 4 ℃ for 24-28h, cleaning, and resuspending to obtain the final product;
(4) growth of gold nanoshell
Mixing carbonate solution and chlorauric acid hydrate (HAuCl4 & 3H)2O) to obtain a colorless solution, then placing the colorless solution at 4 ℃ for aging for 24-28h, then adding the aged solution into the product obtained in the step (3) while stirring, adding a reducing agent after reacting for 8-15min, and continuing stirring until the reduction is complete to obtain the gold nano shell layer.
9. Use according to claim 8, wherein the THPC solution in step (1) is prepared by: 1M of sodium hydroxide, THPC and water are mixed according to the volume ratio of 0.4-0.6: 10-15: 45-50, and mixing.
10. The application of claim 8, wherein the specific steps in the step (2) are as follows: mixing 2.5 wt% of silicon dioxide nanospheres with ethanol, uniformly dispersing by ultrasonic, adding 3-aminopropyl triethoxysilane, carrying out heat treatment at 90 ℃ for 3h, cooling to room temperature, centrifuging, washing with ethanol, and finally carrying out heavy suspension with ethanol to obtain the silicon dioxide nanospheres.
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Cited By (4)
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| CN115491410A (en) * | 2022-10-18 | 2022-12-20 | 浙江大学 | Application of titanium dioxide/zinc-aluminum hydrotalcite oxide nano material in oligonucleotide sequencing |
| CN117074503A (en) * | 2023-10-16 | 2023-11-17 | 成都泰莱医学检验实验室有限公司 | Nanocomposite for time-of-flight mass spectrometry detection and preparation method thereof |
| CN117066505A (en) * | 2023-07-31 | 2023-11-17 | 广东省农业科学院农业生物基因研究中心 | Laser desorption/ionization mass spectrometry method for detecting vitamins |
| CN117092199A (en) * | 2023-10-16 | 2023-11-21 | 成都泰莱医学检验实验室有限公司 | Method for performing time-of-flight mass spectrometry detection by using tungsten carbide nanomaterial matrix |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115491410A (en) * | 2022-10-18 | 2022-12-20 | 浙江大学 | Application of titanium dioxide/zinc-aluminum hydrotalcite oxide nano material in oligonucleotide sequencing |
| CN115491410B (en) * | 2022-10-18 | 2024-05-17 | 浙江大学 | Application of titanium dioxide/zinc-aluminum hydrotalcite like oxide nano material in oligonucleotide sequencing |
| CN117066505A (en) * | 2023-07-31 | 2023-11-17 | 广东省农业科学院农业生物基因研究中心 | Laser desorption/ionization mass spectrometry method for detecting vitamins |
| CN117066505B (en) * | 2023-07-31 | 2024-03-19 | 广东省农业科学院农业生物基因研究中心 | Laser desorption/ionization mass spectrometry method for detecting vitamins |
| CN117074503A (en) * | 2023-10-16 | 2023-11-17 | 成都泰莱医学检验实验室有限公司 | Nanocomposite for time-of-flight mass spectrometry detection and preparation method thereof |
| CN117092199A (en) * | 2023-10-16 | 2023-11-21 | 成都泰莱医学检验实验室有限公司 | Method for performing time-of-flight mass spectrometry detection by using tungsten carbide nanomaterial matrix |
| CN117074503B (en) * | 2023-10-16 | 2024-01-26 | 成都泰莱医学检验实验室有限公司 | Nanocomposite for time-of-flight mass spectrometry detection and preparation method thereof |
| CN117092199B (en) * | 2023-10-16 | 2024-01-26 | 成都泰莱医学检验实验室有限公司 | Method for performing time-of-flight mass spectrometry detection by using tungsten carbide nanomaterial matrix |
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Application publication date: 20200714 |