CN111337462A - Silver nanoparticle fluorescent switch system, preparation method thereof and application thereof in drug detection - Google Patents
Silver nanoparticle fluorescent switch system, preparation method thereof and application thereof in drug detection Download PDFInfo
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- CN111337462A CN111337462A CN202010111942.2A CN202010111942A CN111337462A CN 111337462 A CN111337462 A CN 111337462A CN 202010111942 A CN202010111942 A CN 202010111942A CN 111337462 A CN111337462 A CN 111337462A
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- 239000002105 nanoparticle Substances 0.000 title claims abstract description 63
- 229910052709 silver Inorganic materials 0.000 title claims abstract description 51
- 239000004332 silver Substances 0.000 title claims abstract description 51
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 title claims abstract description 37
- 229940079593 drug Drugs 0.000 title claims abstract description 30
- 239000003814 drug Substances 0.000 title claims abstract description 30
- 238000001514 detection method Methods 0.000 title claims abstract description 26
- 238000002360 preparation method Methods 0.000 title abstract description 19
- MTPVUVINMAGMJL-UHFFFAOYSA-N trimethyl(1,1,2,2,2-pentafluoroethyl)silane Chemical compound C[Si](C)(C)C(F)(F)C(F)(F)F MTPVUVINMAGMJL-UHFFFAOYSA-N 0.000 claims description 50
- FOIXSVOLVBLSDH-UHFFFAOYSA-N Silver ion Chemical compound [Ag+] FOIXSVOLVBLSDH-UHFFFAOYSA-N 0.000 claims description 49
- CWQXQMHSOZUFJS-UHFFFAOYSA-N molybdenum disulfide Chemical compound S=[Mo]=S CWQXQMHSOZUFJS-UHFFFAOYSA-N 0.000 claims description 39
- 229910052982 molybdenum disulfide Inorganic materials 0.000 claims description 39
- MYWUZJCMWCOHBA-VIFPVBQESA-N methamphetamine Chemical compound CN[C@@H](C)CC1=CC=CC=C1 MYWUZJCMWCOHBA-VIFPVBQESA-N 0.000 claims description 37
- 229960001252 methamphetamine Drugs 0.000 claims description 37
- 238000006243 chemical reaction Methods 0.000 claims description 35
- 229910021642 ultra pure water Inorganic materials 0.000 claims description 25
- 239000012498 ultrapure water Substances 0.000 claims description 25
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- YGYXSAHZJOICHH-UHFFFAOYSA-N [Ag].[Mo](=S)=S Chemical compound [Ag].[Mo](=S)=S YGYXSAHZJOICHH-UHFFFAOYSA-N 0.000 claims description 10
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- 238000000034 method Methods 0.000 claims description 10
- SQGYOTSLMSWVJD-UHFFFAOYSA-N silver(1+) nitrate Chemical compound [Ag+].[O-]N(=O)=O SQGYOTSLMSWVJD-UHFFFAOYSA-N 0.000 claims description 10
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 9
- 238000001816 cooling Methods 0.000 claims description 9
- 238000002189 fluorescence spectrum Methods 0.000 claims description 9
- ZOKXTWBITQBERF-UHFFFAOYSA-N Molybdenum Chemical compound [Mo] ZOKXTWBITQBERF-UHFFFAOYSA-N 0.000 claims description 8
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- 239000011733 molybdenum Substances 0.000 claims description 8
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- 238000003756 stirring Methods 0.000 claims description 8
- 229910052717 sulfur Inorganic materials 0.000 claims description 8
- 239000011593 sulfur Substances 0.000 claims description 8
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 claims description 7
- IARLBMZSSJJOCU-UHFFFAOYSA-N sodium;1-oxidopyrrolidine-2,5-dione Chemical compound [Na+].[O-]N1C(=O)CCC1=O IARLBMZSSJJOCU-UHFFFAOYSA-N 0.000 claims description 7
- 238000005406 washing Methods 0.000 claims description 7
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- 239000003446 ligand Substances 0.000 claims description 5
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- 229910001961 silver nitrate Inorganic materials 0.000 claims description 5
- 238000009210 therapy by ultrasound Methods 0.000 claims description 5
- 238000001035 drying Methods 0.000 claims description 4
- 229910009818 Ti3AlC2 Inorganic materials 0.000 claims description 2
- 238000005411 Van der Waals force Methods 0.000 claims description 2
- 238000007865 diluting Methods 0.000 claims description 2
- 238000010438 heat treatment Methods 0.000 claims description 2
- 239000002994 raw material Substances 0.000 claims description 2
- 150000003378 silver Chemical class 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims 1
- 230000035945 sensitivity Effects 0.000 abstract description 7
- 238000002866 fluorescence resonance energy transfer Methods 0.000 abstract description 6
- 239000000126 substance Substances 0.000 description 14
- 230000000052 comparative effect Effects 0.000 description 8
- UMGDCJDMYOKAJW-UHFFFAOYSA-N thiourea group Chemical group NC(=S)N UMGDCJDMYOKAJW-UHFFFAOYSA-N 0.000 description 8
- 230000003993 interaction Effects 0.000 description 7
- 239000000523 sample Substances 0.000 description 7
- 238000001917 fluorescence detection Methods 0.000 description 6
- 239000007850 fluorescent dye Substances 0.000 description 6
- 238000005457 optimization Methods 0.000 description 6
- 230000005284 excitation Effects 0.000 description 5
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 4
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Natural products NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 4
- 238000010521 absorption reaction Methods 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 230000004044 response Effects 0.000 description 4
- 239000001509 sodium citrate Substances 0.000 description 4
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 4
- 239000011684 sodium molybdate Substances 0.000 description 4
- 235000015393 sodium molybdate Nutrition 0.000 description 4
- TVXXNOYZHKPKGW-UHFFFAOYSA-N sodium molybdate (anhydrous) Chemical compound [Na+].[Na+].[O-][Mo]([O-])(=O)=O TVXXNOYZHKPKGW-UHFFFAOYSA-N 0.000 description 4
- 239000012467 final product Substances 0.000 description 3
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- 239000010936 titanium Substances 0.000 description 3
- WHNWPMSKXPGLAX-UHFFFAOYSA-N N-Vinyl-2-pyrrolidone Chemical compound C=CN1CCCC1=O WHNWPMSKXPGLAX-UHFFFAOYSA-N 0.000 description 2
- 239000004809 Teflon Substances 0.000 description 2
- 229920006362 Teflon® Polymers 0.000 description 2
- 239000011609 ammonium molybdate Substances 0.000 description 2
- 235000018660 ammonium molybdate Nutrition 0.000 description 2
- APUPEJJSWDHEBO-UHFFFAOYSA-P ammonium molybdate Chemical group [NH4+].[NH4+].[O-][Mo]([O-])(=O)=O APUPEJJSWDHEBO-UHFFFAOYSA-P 0.000 description 2
- 229940010552 ammonium molybdate Drugs 0.000 description 2
- 229960005070 ascorbic acid Drugs 0.000 description 2
- 235000010323 ascorbic acid Nutrition 0.000 description 2
- 239000011668 ascorbic acid Substances 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
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- 238000010791 quenching Methods 0.000 description 2
- 230000000171 quenching effect Effects 0.000 description 2
- 229910052979 sodium sulfide Inorganic materials 0.000 description 2
- GRVFOGOEDUUMBP-UHFFFAOYSA-N sodium sulfide (anhydrous) Chemical compound [Na+].[Na+].[S-2] GRVFOGOEDUUMBP-UHFFFAOYSA-N 0.000 description 2
- YUKQRDCYNOVPGJ-UHFFFAOYSA-N thioacetamide Chemical compound CC(N)=S YUKQRDCYNOVPGJ-UHFFFAOYSA-N 0.000 description 2
- DLFVBJFMPXGRIB-UHFFFAOYSA-N thioacetamide Natural products CC(N)=O DLFVBJFMPXGRIB-UHFFFAOYSA-N 0.000 description 2
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- 239000011541 reaction mixture Substances 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 229940043263 traditional drug Drugs 0.000 description 1
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
- G01N21/643—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" non-biological material
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/33—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using ultraviolet light
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- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N2021/6417—Spectrofluorimetric devices
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
- G01N2021/6432—Quenching
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Abstract
The invention relates to a silver nanoparticle fluorescence switch system, a preparation method thereof and application thereof in drug detection. Compared with the prior art, the fluorescence resonance energy transfer system for detecting the drugs is successfully constructed by selecting the proper donor molecules and the proper acceptor molecules, and when the system is used for detecting the drugs, the background interference is smaller, the reliability is higher, and the sensitivity and the selectivity are higher.
Description
Technical Field
The invention belongs to the field of drug detection, and particularly relates to a silver nanoparticle fluorescent switch system, a preparation method thereof and application thereof in drug detection.
Background
At present, the field of drug detection is mainly instrument detection, but has some disadvantages, such as: expensive instrument, complex operation, long pretreatment time, inconvenient field detection and the like. Fluorescence detection has certain advantages, such as: simple, convenient, fast, economical, portable and the like. The general fluorescence detection is mostly used for detecting analytes in a fluorescence quenching mode, and the mode is influenced by too many external factors, so that the detection result is not high enough in accuracy. And a fluorescence switch type detection probe is constructed based on fluorescence resonance energy transfer, and an analyte is detected in a fluorescence enhanced mode, so that the background interference is less, the reliability is higher, and the sensitivity and the selectivity are higher. At present, a fluorescence switch type detection probe is constructed based on a fluorescence resonance energy transfer system, and reports of the fluorescence switch type detection probe for the field of drug detection are few.
When the fluorescence spectrum of one fluorescent molecule (also called donor molecule) overlaps with the absorption spectrum of another molecule (also called acceptor molecule), the excitation energy of the donor fluorescent molecule induces the acceptor molecule to emit fluorescence, and the fluorescence intensity of the donor fluorescent molecule is attenuated to form a fluorescence resonance energy transfer system. In the construction of a fluorescence resonance energy transfer system, the conditions under which efficient energy transfer occurs between the energy donor-acceptor (D-a) pair are harsh, and mainly include: (1) the emission spectrum of the energy donor must overlap with the absorption spectrum of the energy acceptor; (2) the fluorescent chromophores of the energy donor and the energy acceptor must be arranged in a suitable manner; (3) the energy donor and the energy acceptor must be close enough to each other so that the probability of energy transfer is high. In addition, there are many requirements for suitable donor and acceptor molecules in terms of quantum yield, extinction coefficient, water solubility, interference resistance, etc. It can be seen that finding a suitable D-A pair is difficult.
Disclosure of Invention
The invention aims to solve the technical problems that when a fluorescence quenching mode is used for detecting drugs in the traditional drug detection field, external interference factors are more, and the detection sensitivity and result accuracy are not high enough, and provides a silver nanoparticle fluorescence switch system, a preparation method thereof and application thereof in drug detection.
The invention realizes the purpose through the following technical scheme:
the invention provides a silver nanoparticle fluorescence switch system for detecting the content of drugs, which comprises silver nanoparticles and fluorescence donor molecules, wherein the silver nanoparticles are modified with antibodies capable of being specifically combined with the drugs to be detected on the surfaces, and the fluorescence emission spectrum of the fluorescence donor molecules is overlapped with the ultraviolet visible absorption spectrum of the silver nanoparticles.
As a further optimization scheme of the invention, the fluorescence donor molecule is fluorescence molybdenum disulfide to form a molybdenum disulfide-silver nanoparticle fluorescence switch system; the fluorescence emission spectrum of the molybdenum disulfide and the ultraviolet visible absorption spectrum of the silver nanoparticles are overlapped, when the two are mixed, due to the characteristics of the two-dimensional material of the molybdenum disulfide, the molybdenum disulfide has the characteristics of large specific surface area and easy adsorption, the molybdenum disulfide and the silver nanoparticles are mutually connected together through van der Waals force of the surfaces, and the fluorescence of the system is quenched at the moment; when the drug to be detected exists, the silver nanoparticles are modified on the surfaces thereof with corresponding antibodies and can be combined with the antibodies, and the combination force is stronger than the interaction between the silver nanoparticles and molybdenum disulfide, so that the silver nanoparticles are far away from the molybdenum disulfide, and the fluorescence of the system is recovered.
As a further optimization scheme of the invention, the fluorescence donor molecule is fluorescent titanium carbide MXene to form a titanium carbide MXene-silver nanoparticle fluorescence switch system; the fluorescence emission spectrum of the titanium carbide MXene and the ultraviolet-visible absorption spectrum of the silver nanoparticles are overlapped, when the titanium carbide MXene and the silver nanoparticles are mixed, the titanium carbide MXene surface contains rich functional groups and a large number of hydrophilic groups and can be connected with the surface functional groups of the silver nanoparticles through the interaction of hydrogen bonds and the like, and the fluorescence of the system is quenched; when the drug to be detected exists, the silver nanoparticles are modified on the surfaces thereof with corresponding antibodies and can be combined with the antibody, and the combination force is stronger than the interaction between the silver nanoparticles and the titanium carbide MXene, so that the silver nanoparticles are far away from the titanium carbide MXene, and the fluorescence of the system is recovered.
The invention also provides a preparation method of the silver nanoparticle fluorescent switch system, which comprises the following steps:
(1) preparing a fluorescence donor molecule;
(2) preparing silver nanoparticles;
(3) carrying out antibody modification on the surface of the silver nano particles by utilizing an antibody specifically bound with the drug to be detected;
(4) preparing a fluorescence donor molecule-silver nanoparticle composite material: and mixing the fluorescence donor molecules with the silver nanoparticles modified by the antibody to obtain the fluorescence switch system.
As a further optimization scheme of the invention, the fluorescence donor molecule is fluorescence molybdenum disulfide, and the preparation method of the fluorescence molybdenum disulfide comprises the following steps: taking a molybdenum source and a sulfur source, and according to the sulfur source: the molar ratio of the molybdenum source is 5: 1-2: mixing the raw materials in the ultrapure water according to the proportion of 1, adjusting the pH value to 1, reacting in a polytetrafluoroethylene high-pressure kettle at the temperature of 180 ℃ and 240 ℃ for 18-24h, cooling and centrifuging after the reaction is finished, collecting a product, and drying for later use; the molybdenum source is selected from ammonium molybdate or sodium molybdate and the like, and the sulfur source is selected from thiourea, thioacetamide, sodium sulfide and the like.
As a further optimization scheme of the invention, the fluorescence donor molecule is fluorescence titanium carbide MXene, and the preparation method of the fluorescence titanium carbide MXene comprises the following steps: mixing concentrated hydrochloric acid and lithium fluoride in ultrapure water, stirring, and adding Ti3AlC2Mixing the powder evenly, and reacting for 18-24h at 25-50 ℃; after the reaction is finished, centrifuging, washing with water until the pH of the supernatant is 5.0-6.5, and collecting to obtain an intermediate product A; dissolving the intermediate product A in ultrapure water, introducing inert gas to remove oxygen, performing ultrasonic treatment, centrifuging, and collecting supernatant to obtain an intermediate product B; diluting the intermediate product B, dispersing the intermediate product B in ethanol or dimethylformamide, introducing inert gas to remove oxygen, then placing the intermediate product B in a polytetrafluoroethylene high-pressure reaction kettle, and reacting for 6-18h at the temperature of 110-; and after the reaction is finished, filtering and centrifuging, and collecting supernatant to obtain a product for later use.
As a further optimized scheme of the invention, the method for preparing the silver nanoparticles comprises the following steps: dispersing silver nitrate in ultrapure water, heating to 100 ℃, adding a ligand for reaction, obtaining a product after the reaction is finished, and cooling for later use; the ligand is sodium citrate, PVP or ascorbic acid, etc.
As a further optimization scheme of the invention, the method for performing antibody modification on the surface of the silver nanoparticle comprises the following steps: adding 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide sodium salt into the prepared silver nanoparticle solution, mixing and stirring at room temperature, adding a monoclonal antibody specifically bound with the drug to be tested, and incubating to obtain the antibody modified silver nanoparticle solution.
The invention also provides an application of the silver nanoparticle fluorescent switch system in drug detection.
As a further optimization scheme of the invention, the drug is methamphetamine.
The invention has the beneficial effects that: the invention provides a silver nanoparticle fluorescence switch system, a preparation method thereof and application in drug detection, wherein a fluorescence resonance energy transfer system for detecting drugs is successfully constructed by selecting proper donor molecules and acceptor molecules, and when drugs are detected by using the system, the background interference is smaller, the reliability is higher, and the sensitivity and the selectivity are higher.
Drawings
FIG. 1 is a graph of the fluorescent response of the molybdenum disulfide-silver nanoparticle fluorescent switch system of example 1 to methamphetamine;
FIG. 2 is a graph of the linear relationship of the molybdenum disulfide-silver nanoparticle fluorescent switch system of example 1 to the content of methamphetamine;
FIG. 3 is a graph of the fluorescence response of the titanium carbide MXene-silver nanoparticle fluorescent switch system of example 2 to methamphetamine;
FIG. 4 is a graph of the linear relationship of the titanium carbide MXene-silver nanoparticle fluorescent switch system of example 2 to the content of methamphetamine;
FIG. 5 is a graph of the fluorescent response of a molybdenum disulfide fluorescent probe to methamphetamine in comparative example 1;
FIG. 6 is a graph of the linear relationship of molybdenum disulfide fluorescence probe to methamphetamine content of comparative example 1;
FIG. 7 is a graph showing the fluorescence response of a titanium carbide MXene fluorescent probe to methamphetamine in comparative example 2;
FIG. 8 is a linear relationship graph of the content of methamphetamine in the titanium carbide MXene fluorescent probe of comparative example 2.
Detailed Description
The present invention is described in detail below with reference to specific embodiments, it should be noted that the following specific embodiments are only used for further description of the present application, and should not be construed as limiting the scope of the present application, and those skilled in the art can make modifications and adaptations of the present application based on the above-mentioned content.
The preparation method of the silver nanoparticle fluorescent switch system comprises the following steps:
(1) preparing a fluorescence donor molecule, including preparing fluorescence molybdenum disulfide and preparing fluorescence titanium carbide MXene:
preparation of fluorescent molybdenum disulfide: taking a molybdenum source and a sulfur source, and according to the sulfur source: the molar ratio of the molybdenum source is 5: 1-2: 1 proportion is mixed in ultrapure water, the pH value is adjusted to 1, the mixture is reacted in a polytetrafluoroethylene high-pressure kettle for 18 to 24 hours at the temperature of 180 ℃ and 240 ℃, after the reaction is finished, the mixture is cooled and centrifuged, the product is collected, and the fluorescent molybdenum disulfide is obtained and dried for standby application; the molybdenum source is selected from ammonium molybdate or sodium molybdate and the like, and the sulfur source is selected from thiourea, thioacetamide or sodium sulfide and the like.
Preparation of fluorescent titanium carbide MXene: 2ml of concentrated hydrochloric acid and 0.3g of lithium fluoride in 5ml of ultrapure water were mixed and stirred, and uniformly dispersed. 0.5g of Ti was further added3AlC2Mixing the powder and reacting at 25-50 deg.c for 18-24 hr. After the reaction is finished, centrifuging, washing with water until the pH of the supernatant is about 5.0-6.5, and collecting an intermediate product A. Dissolving A in 3ml of ultrapure water, introducing inert gas to remove oxygen, performing ultrasonic treatment for 3-5 hours, performing low-speed centrifugation (about 2500-. Taking 2ml of the product B, dispersing the product B in 20ml of ethanol or dimethylformamide, introducing inert gas to remove oxygen, and reacting for 6-18 hours at the temperature of 110-150 ℃ in a polytetrafluoroethylene stainless steel reaction kettle. After the reaction is finished, filtering and precipitating by using a 0.22 mu m microporous filtering membrane, centrifuging the upper layer solution at high speed again (8000- & ltSUB & gt 12000 r/min), and collecting the supernatant to obtain a final product C, namely the fluorescent titanium carbide solution.
(2) Preparation of silver nanoparticles
100-120mg of silver nitrate and 200ml of ultrapure water are uniformly dispersed; the reaction mixture was heated to 100 ℃ and 0.1g of ligand was added rapidly and reacted for 1 hour. And after the reaction is finished, obtaining silver nanoparticle solution, and cooling for later use. The ligand is sodium citrate, PVP or ascorbic acid, etc., but is not limited thereto.
(3) Antibody modification of silver nanoparticle surface by using antibody specifically bound with drug to be detected
Take 0.5 × 10-5mmol-2×10-5mmol of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride, 0.5 × 10-5mmol-2×10-5Adding mmol N-hydroxysuccinimide sodium salt into silver nanoparticle solution, mixing and stirring at room temperature for 15 min-4 hr, adding 0.1 × 10-5mmol-5×10-5And (3) incubating the mmol monoclonal antibody capable of being specifically bound with the drug to be detected for 1-6 hours to obtain the silver nanoparticle solution of the modified antibody.
(4) Preparing a fluorescence donor molecule-silver nanoparticle composite material:
preparing a molybdenum disulfide-silver nanoparticle fluorescent switch system: dispersing 10mg of fluorescent molybdenum disulfide in 10ml of ultrapure water, adding 0.2ml to 2ml of the silver nanoparticle solution of the modified antibody, and uniformly mixing.
Preparing a titanium carbide MXene-silver nanoparticle fluorescent switch system: and (3) adding 10ml of fluorescent titanium carbide MXene into 0.2ml to 2ml of the silver nanoparticle solution of the modified antibody, and uniformly mixing.
The present application is described in further detail below by way of specific examples.
Example 1
The embodiment provides a preparation method of a molybdenum disulfide-silver nanoparticle fluorescent switch system for detecting methamphetamine, which comprises the following steps:
(1) preparation of fluorescent molybdenum disulfide: 5mmol of sodium molybdate, 10mmol of thiourea and 50ml of ultrapure water are mixed and stirred uniformly; the pH was adjusted to 1, and the reaction was carried out in a polytetrafluoroethylene autoclave at 200 ℃ for 20 hours. After the reaction is finished, cooling, centrifuging, washing, collecting the product, and drying at 40 ℃ for later use.
(2) Preparing silver nanoparticles: 100mg of silver nitrate and 200ml of ultrapure water are uniformly dispersed; the mixture was heated to 100 ℃ and 0.1g of sodium citrate was added rapidly to react for 1 hour. And after the reaction is finished, cooling for later use. The synthesized silver nanoparticles are about 30nm, the absorption peak of the silver nanoparticles is about 405nm, and the absorption spectrum of the silver nanoparticles is overlapped with the fluorescence emission spectrum of the fluorescent molybdenum disulfide.
(3) Silver nanoparticle surface antibody modification 1 × 10-5mmol of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride, 1 × 10-5mmol N-hydroxysuccinimide sodium salt, 10ml silver nano-particle solution, mixing and stirring for 1 hour at room temperature, then adding 1 × 10-5mmol of methamphetamine monoclonal antibody was incubated at 37 ℃ for 4 hours.
(4) Preparing a molybdenum disulfide-silver nanoparticle composite material: dispersing 10mg of fluorescent molybdenum disulfide in 10ml of ultrapure water, adding 1ml of the silver nanoparticle solution with the modified antibody, and uniformly mixing.
Detection of methamphetamine by the system: under excitation of 255nm, the fluorescent molybdenum disulfide has the strongest fluorescence emission at 405 nm. The absorption peak of the silver nanoparticles is positioned at 403nm, and the ultraviolet visible absorption spectrum after the antibody is modified is basically unchanged. Because the fluorescence emission spectrum of the molybdenum disulfide is overlapped with the ultraviolet visible absorption spectrum of the silver nanoparticles, when the molybdenum disulfide and the silver nanoparticles are mixed, the molybdenum disulfide and the silver nanoparticles are connected together through weak interaction of the surfaces due to the characteristics of two-dimensional materials of the molybdenum disulfide, and the fluorescence of the system is quenched. When methamphetamine exists, the silver nanoparticles are modified on the surfaces thereof with corresponding antibodies and can be combined with the antibodies, the combination force is stronger than the interaction between the silver nanoparticles and molybdenum disulfide, therefore, the silver nanoparticles are far away from the molybdenum disulfide, the fluorescence of the system is recovered, and the target object is detected in a fluorescence switch mode. And (3) plotting by taking the ratio of the intensity variation of the fluorescence emission peak at 405nm to the fluorescence intensity without adding the substance to be detected as the ordinate when the substance to be detected is added and the substance to be detected is not added and the molar concentration of the methamphetamine as the abscissa to obtain linear correlation, and taking the linear correlation as the fluorescence detection of the methamphetamine with unknown content.
Example 2
The embodiment provides a preparation method of a titanium carbide MXene-silver nanoparticle fluorescent switch system for detecting methamphetamine, which comprises the following steps:
(1) preparation of fluorescent titanium carbide MXene: 2ml of concentrated hydrochloric acid and 0.3g of lithium fluoride in 5ml of ultrapure water were mixed and stirred, and uniformly dispersed. 0.5g of Ti was further added3AlC2The powders are mixed evenly and reacted for 20 hours at 40 ℃. After the reaction is finished, centrifuging, washing with water until the pH of the supernatant is about 6, and collecting an intermediate product A. Dissolving A in 3ml of ultrapure water, N2Bubbling for 30 minutes, carrying out ultrasonic treatment for 4 hours, centrifuging once (4000 rpm), collecting supernatant, and metering to 10ml (marked as intermediate product B) by using ultrapure water for standby. 2ml of product B are taken and dispersed in 20ml of ethanol, N2Bubbling was carried out for 30 minutes, and the reaction was carried out in a Teflon stainless steel reaction vessel at 120 ℃ for 12 hours. After the reaction is finished, filtering and precipitating by using a 0.22-micron microfiltration membrane, centrifuging the upper-layer solution again (9000 r/min), and collecting the supernatant to obtain a final product C, namely the fluorescent titanium carbide MXene solution.
(2) Preparing silver nanoparticles: 120mg of silver nitrate and 200ml of ultrapure water are uniformly dispersed; the mixture was heated to 100 ℃ and 0.1g of sodium citrate was added rapidly to react for 1 hour. And after the reaction is finished, cooling for later use. The synthesized silver nanoparticles are about 35nm, so that the absorption peak of the silver nanoparticles is about 410nm and is overlapped with the fluorescence emission spectrum of the fluorescence titanium carbide MXene.
(3) Silver nanoparticle surface antibody modification 1 × 10-5mmol of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride, 1 × 10-5mmol N-hydroxysuccinimide sodium salt, 10ml silver nano-particle solution, mixing and stirring for 1 hour at room temperature, then adding 1 × 10-5mmol of methamphetamine monoclonal antibody was incubated at 37 ℃ for 4 hours.
(4) Preparing a titanium carbide MXene-silver nanoparticle composite material: and (3) adding 10ml of the prepared fluorescent titanium carbide MXene into 1ml of the silver nanoparticle solution of the modified antibody, and uniformly mixing.
Detection of methamphetamine by the system: under 350nm excitation, the fluorescent titanium carbide MXene has the strongest fluorescence emission at 450 nm. The absorption peak of the silver nanoparticles is positioned at 410nm, and the ultraviolet visible absorption spectrum after the antibody is modified is basically unchanged. The fluorescence emission spectrum of the titanium carbide MXene is overlapped with the ultraviolet visible absorption spectrum of the silver nanoparticles, and the titanium carbide MXene surface contains abundant functional groups and a large number of hydrophilic groups and can be connected with the surface functional groups of the silver nanoparticles through interaction such as hydrogen bonds, so when the titanium carbide MXene and the silver nanoparticles are mixed, the fluorescence of the system is quenched. When methamphetamine exists, the silver nanoparticles are modified on the surfaces thereof with corresponding antibodies and can be combined with the antibodies, the combination force is stronger than the interaction between the silver nanoparticles and the titanium carbide MXene, therefore, the silver nanoparticles are far away from the titanium carbide MXene, the fluorescence of the system is recovered, and the target object is detected by means of the fluorescence switch. And (3) when the substance to be detected is added and the substance to be detected is not added, the ratio of the fluorescence emission peak intensity variation at 450nm to the fluorescence intensity when the substance to be detected is not added is taken as the ordinate, the molar concentration of the methamphetamine is taken as the abscissa, and the linear correlation is obtained by plotting and taken as the fluorescence detection of the methamphetamine with unknown content.
Comparative example 1
The comparative example constructs a molybdenum disulfide fluorescent probe for detecting methamphetamine, and the steps comprise:
(1) preparation of fluorescent molybdenum disulfide: 5mmol of sodium molybdate, 10mmol of thiourea and 50ml of ultrapure water are mixed and stirred uniformly; the pH was adjusted to 1, and the reaction was carried out in a polytetrafluoroethylene autoclave at 200 ℃ for 20 hours. After the reaction is finished, cooling, centrifuging, washing, collecting the product, and drying at 40 ℃ for later use.
(2) Fluorescent molybdenum disulfide surface antibody modification 1 × 10-5mmol of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride, 1 × 10-5mmol N-hydroxysuccinimide sodium salt, 10mg molybdenum disulfide, in 10ml ultrapure water, at room temperature, mixing and stirring for 1 hour, then adding 1 × 10-5mmol of methamphetamine monoclonal antibody was incubated at 37 ℃ for 4 hours.
Detection of methamphetamine by the probe: under excitation of 255nm, the fluorescent molybdenum disulfide has the strongest fluorescence emission at 405 nm. When the substance to be detected exists, the fluorescence intensity of the molybdenum disulfide is reduced. And (3) when the substance to be detected is added and the substance to be detected is not added, the ratio of the intensity variation of the fluorescence emission peak at 405nm to the fluorescence intensity when the substance to be detected is not added is used as the ordinate, the molar concentration of the methamphetamine is used as the abscissa, and the linear correlation is obtained by plotting and is used as the fluorescence detection of the methamphetamine with unknown content.
Comparative example 2
The comparative example constructs a titanium carbide MXene fluorescent probe for detecting methamphetamine, and the steps comprise:
(1) preparation of fluorescent titanium carbide MXene: 2ml of concentrated hydrochloric acid and 0.3g of lithium fluoride in 5ml of ultrapure water were mixed and stirred, and uniformly dispersed. 0.5g of Ti was further added3AlC2The powders are mixed evenly and reacted for 20 hours at 40 ℃. After the reaction is finished, centrifuging, washing with water until the pH of the supernatant is about 6, and collecting an intermediate product A. Dissolving A in 3ml of ultrapure water, N2Bubbling for 30 minutes, carrying out ultrasonic treatment for 4 hours, centrifuging once (4000 rpm), collecting supernatant, and metering to 10ml (marked as intermediate product B) by using ultrapure water for standby. 2ml of product B are taken and dispersed in 20ml of ethanol, N2Bubbling was carried out for 30 minutes, and the reaction was carried out in a Teflon stainless steel reaction vessel at 120 ℃ for 12 hours. After the reaction is finished, filtering and precipitating by using a 0.22-micron microfiltration membrane, centrifuging the upper-layer solution again (9000 r/min), and collecting the supernatant to obtain a final product C, namely the fluorescent titanium carbide MXene solution.
(2) Titanium carbide MXene surface antibody modification 1 × 10-5mmol of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride, 1 × 10-5mmol of N-hydroxysuccinimide sodium salt and 10ml of the above titanium carbide MXene solution were mixed and stirred at room temperature for 1 hour, and 1 × 10 was added-5mmol of methamphetamine monoclonal antibody was incubated at 37 ℃ for 4 hours.
Detection of methamphetamine by the probe: under 350nm excitation, titanium carbide MXene has the strongest fluorescence emission at 450 nm. When the substance to be detected exists, the fluorescence intensity of the titanium carbide MXene is reduced. And (3) when the substance to be detected is added and the substance to be detected is not added, the ratio of the fluorescence emission peak intensity variation at 450nm to the fluorescence intensity when the substance to be detected is not added is taken as the ordinate, the molar concentration of the methamphetamine is taken as the abscissa, and the linear correlation is obtained by plotting and taken as the fluorescence detection of the methamphetamine with unknown content.
The results are shown in FIGS. 1-8, which show that the fluorescent probe of the invention has stable intensity and higher detection sensitivity to methamphetamine. As shown in fig. 2, 4, 6, and 8, the conversion formula of the molybdenum disulfide-silver nanoparticle system to the standard concentration of methamphetamine is: y 3.0812+0.3288x, R20.9913. The conversion formula of the standard concentration of the methamphetamine detected by the titanium carbide MXene-silver nanoparticle system is as follows: y 5.2916+0.5701x, R20.9920. The conversion formula of the molybdenum disulfide to the standard concentration of methamphetamine detection is as follows: y 0.8271+0.1247x, R20.9901, the conversion formula of the standard concentration of the methamphetamine detected by the titanium carbide MXene is as follows: y 1.2587+0.1799x, R20.9903. The slope in the linear relation between the titanium carbide MXene-silver nanoparticle system and the molybdenum disulfide-silver nanoparticle system is obviously greater than that of the titanium carbide MXene and the molybdenum disulfide system, wherein the slope of the titanium carbide MXene-silver nanoparticle system is greater than that of the molybdenum disulfide-silver nanoparticle system, which shows that under an object to be detected with the same concentration, a fluorescence switch detection probe constructed by the titanium carbide MXene or the molybdenum disulfide and the silver nanoparticles has the advantages of larger signal change, higher signal-to-noise ratio and higher sensitivity compared with the titanium carbide MXene or the molybdenum disulfide. In particular, the titanium carbide MXene-silver nanoparticle system has larger signal change and higher sensitivity.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the present invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention.
Claims (10)
1. A silver nanoparticle fluorescence switch system is used for detecting the content of drugs and is characterized by comprising silver nanoparticles and fluorescence donor molecules, wherein the surfaces of the silver nanoparticles are modified with antibodies specifically bound with the drugs to be detected, and the fluorescence emission spectrum of the fluorescence donor molecules is overlapped with the ultraviolet visible absorption spectrum of the silver nanoparticles.
2. The silver nanoparticle fluorescent switch system according to claim 1, wherein the fluorescent donor molecule is fluorescent molybdenum disulfide, and the silver nanoparticle is adsorbed on the surface of the fluorescent molybdenum disulfide by van der waals force to form a molybdenum disulfide-silver nanoparticle fluorescent switch system.
3. The silver nanoparticle fluorescent switch system according to claim 1, wherein the fluorescence donor molecule is fluorescent titanium carbide MXene, and the silver nanoparticles are connected with functional groups and/or hydrophilic groups on the surface of the titanium carbide MXene through surface functional groups to form a titanium carbide MXene-silver nanoparticle fluorescent switch system.
4. A method of making a silver nanoparticle fluorescent switching system as claimed in any one of claims 1 to 3, comprising the steps of:
(1) preparing a fluorescence donor molecule;
(2) preparing silver nanoparticles;
(3) carrying out antibody modification on the surface of the silver nano particles by utilizing an antibody specifically bound with the drug to be detected;
(4) preparing a fluorescence donor molecule-silver nanoparticle composite material: and mixing the fluorescence donor molecules with the silver nanoparticles modified by the antibody to obtain the fluorescence switch system.
5. The method for preparing a silver nanoparticle fluorescent switch system according to claim 4, wherein the fluorescent donor molecule is fluorescent molybdenum disulfide, and the method for preparing the fluorescent molybdenum disulfide comprises the following steps: taking a molybdenum source and a sulfur source, and according to the sulfur source: the molar ratio of the molybdenum source is 5: 1-2: mixing the raw materials in the proportion of 1 in ultrapure water, adjusting the pH value to 1, reacting in a polytetrafluoroethylene autoclave at the temperature of 180 ℃ and 240 ℃ for 18-24h, cooling and centrifuging after the reaction is finished, collecting the product, and drying for later use.
6. The method for preparing the silver nanoparticle fluorescent switch system according to claim 4, wherein the fluorescence donor molecule is fluorescent titanium carbide MXene, and the method for preparing the fluorescent titanium carbide MXene comprises the following steps: mixing concentrated hydrochloric acid and lithium fluoride in ultrapure water, stirring, and adding Ti3AlC2Mixing the powder evenly, and reacting for 18-24h at 25-50 ℃; after the reaction is finished, centrifuging, washing with water until the pH of the supernatant is 5.0-6.5, and collecting to obtain an intermediate product A; dissolving the intermediate product A in ultrapure water, introducing inert gas to remove oxygen, performing ultrasonic treatment, centrifuging, and collecting supernatant to obtain an intermediate product B; diluting the intermediate product B, dispersing the intermediate product B in ethanol or dimethylformamide, introducing inert gas to remove oxygen, then placing the intermediate product B in a polytetrafluoroethylene high-pressure reaction kettle, and reacting for 6-18h at the temperature of 110-; and after the reaction is finished, filtering and centrifuging, and collecting supernatant to obtain a product for later use.
7. The method for preparing a silver nanoparticle fluorescent switch system according to claim 4, wherein the method for preparing the silver nanoparticles comprises the following steps: dispersing silver nitrate in ultrapure water, heating to 100 ℃, adding a ligand for reaction, obtaining a product after the reaction is finished, and cooling for later use.
8. The method for preparing a silver nanoparticle fluorescent switch system according to claim 7, wherein the method for performing antibody modification on the surface of the silver nanoparticle comprises the following steps: adding 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide sodium salt into the prepared silver nanoparticle solution, mixing and stirring at room temperature, adding a monoclonal antibody specifically bound with the drug to be tested, and incubating to obtain the antibody modified silver nanoparticle solution.
9. Use of a silver nanoparticle fluorescent switch system as defined in any one of claims 1 to 3 in drug detection.
10. The use of the silver nanoparticle fluorescent switch system of claim 9, wherein the drug is methamphetamine.
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