CN113358625A - Microneedle patch with plasma enhancement effect and preparation method and application thereof - Google Patents
Microneedle patch with plasma enhancement effect and preparation method and application thereof Download PDFInfo
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- PNEYBMLMFCGWSK-UHFFFAOYSA-N aluminium oxide Inorganic materials [O-2].[O-2].[O-2].[Al+3].[Al+3] PNEYBMLMFCGWSK-UHFFFAOYSA-N 0.000 claims description 3
- 238000010382 chemical cross-linking Methods 0.000 claims description 3
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- 238000006116 polymerization reaction Methods 0.000 claims description 3
- 229910001845 yogo sapphire Inorganic materials 0.000 claims description 3
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- 238000004458 analytical method Methods 0.000 abstract description 2
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- 239000011259 mixed solution Substances 0.000 description 5
- ROOXNKNUYICQNP-UHFFFAOYSA-N ammonium persulfate Chemical compound [NH4+].[NH4+].[O-]S(=O)(=O)OOS([O-])(=O)=O ROOXNKNUYICQNP-UHFFFAOYSA-N 0.000 description 4
- 238000004611 spectroscopical analysis Methods 0.000 description 4
- QPLDLSVMHZLSFG-UHFFFAOYSA-N Copper oxide Chemical compound [Cu]=O QPLDLSVMHZLSFG-UHFFFAOYSA-N 0.000 description 3
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 238000001035 drying Methods 0.000 description 3
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- CYUZOYPRAQASLN-UHFFFAOYSA-N 3-prop-2-enoyloxypropanoic acid Chemical compound OC(=O)CCOC(=O)C=C CYUZOYPRAQASLN-UHFFFAOYSA-N 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
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- 238000004416 surface enhanced Raman spectroscopy Methods 0.000 description 2
- 238000000479 surface-enhanced Raman spectrum Methods 0.000 description 2
- 229910021642 ultra pure water Inorganic materials 0.000 description 2
- 239000012498 ultrapure water Substances 0.000 description 2
- MPNXSZJPSVBLHP-UHFFFAOYSA-N 2-chloro-n-phenylpyridine-3-carboxamide Chemical compound ClC1=NC=CC=C1C(=O)NC1=CC=CC=C1 MPNXSZJPSVBLHP-UHFFFAOYSA-N 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- 206010016952 Food poisoning Diseases 0.000 description 1
- 208000019331 Foodborne disease Diseases 0.000 description 1
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- 229910052709 silver Inorganic materials 0.000 description 1
- 239000004332 silver Substances 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- 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 1
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- RRHXZLALVWBDKH-UHFFFAOYSA-M trimethyl-[2-(2-methylprop-2-enoyloxy)ethyl]azanium;chloride Chemical compound [Cl-].CC(=C)C(=O)OCC[N+](C)(C)C RRHXZLALVWBDKH-UHFFFAOYSA-M 0.000 description 1
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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
- G01N21/65—Raman scattering
- G01N21/658—Raman scattering enhancement Raman, e.g. surface plasmons
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- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Physics & Mathematics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
The invention belongs to the technical field of sample analysis, and particularly relates to a microneedle patch with a plasma enhancement effect, and a preparation method and application thereof. The micro-needle patch with the plasma enhancement effect takes the micro-needle patch as a carrier, and nanoparticles with the plasma resonance effect are loaded on the surface of the micro-needle patch. The microneedle patch can quickly enrich analytes to be detected in a solid sample, enhances Raman detection signals by utilizing the plasma effect of nanoparticles loaded on the surface of the microneedle patch, realizes quick detection of microorganisms and the like in food, and has the characteristics of simple and convenient operation, greatly shortened detection time and the like.
Description
Technical Field
The invention belongs to the technical field of sample analysis, and particularly relates to a microneedle patch with a plasma enhancement effect, and a preparation method and application thereof.
Background
Microorganisms are widely present in nature and in the environment, and in recent years, food safety problems caused by pathogenic microorganisms have been highlighted. According to the statistics of the world health organization, billions of food poisoning events caused by food-borne pathogenic bacteria in every year worldwide seriously threaten human health. Therefore, the method for quickly, accurately and efficiently detecting pathogenic microorganisms is constructed, and has important practical significance.
The Raman spectrum can provide a vibration spectrum of an analyte, so that the material structure information of the analyte is revealed, and the Raman spectrum is suitable for on-site on-line monitoring of pathogenic microorganisms. However, the raman scattering signal generated by the microorganism itself is weak and is easily interfered by autofluorescence, so that there is a certain limitation in the actual detection process. In addition, the pretreatment method for microbial detection is complex and complicated to operate, and takes 3-5 hours. The Surface Enhanced Raman Spectroscopy (SERS) can effectively overcome the defect of Raman scattering, and the key point is how to prepare an active substrate which is efficient and stable and can greatly enhance Raman signals.
Microneedle patches are a new type of carrier, usually made of glass, metal, silica gel or other polymeric materials. Particularly, after the microneedle patch prepared from the polymer with the three-dimensional network is pressed on the surface of a biological tissue, target molecules can be gathered around the tip end of the microneedle through capillary flow driven by swelling, so that the aim of rapid extraction is fulfilled, and the microneedle patch has the characteristics of high efficiency, economy and safety. The method is successfully applied to the aspects of human tissue fluid biological signal extraction and monitoring, plant disease monitoring and the like. Gold, silver, copper and other noble metal nano systems have been widely used in the SERS field due to their excellent optical properties. By utilizing the active substrate loaded with the nano particles, Raman detection signals can be greatly enhanced, and high-sensitivity analysis of a target object to be detected is realized.
Therefore, the adhesion of the microneedle patch is enhanced by ozone treatment, the loading of the nanoparticles on the surface of the needle body is realized, and the microneedle patch with the plasma enhancement effect is expected to be obtained.
Through the current description of the Raman spectrum detection technology and the introduction of the advantages of the microneedle patch and the nanoparticles, the invention provides the microneedle patch with the plasma enhancement effect and the preparation method and the application thereof.
Disclosure of Invention
In view of the above, the present invention provides a microneedle patch with a plasma enhanced effect, and a method for preparing the same and an application thereof.
In order to achieve the purpose, the invention adopts the following technical scheme:
a micro-needle patch with a plasma enhancement effect takes a micro-needle patch as a carrier, and nanoparticles with a plasma resonance effect are loaded on the surface of the micro-needle patch.
The microneedle patch has a surface plasma enhancement effect through surface plasma treatment, and has an amplified signal value when being applied to Raman spectrum detection.
Preferably, the microneedle patch has a chemical crosslinking structure. After the food matrix is inserted, the target to be detected can be quickly enriched by virtue of a swelling effect, and the complete shape and structure can still be kept when the food matrix is stripped from food.
Preferably, the nano-particles are nano-gold, nano-silver, CuO and Al2O3、MgO、Fe2O3、ZnO、ZrO2One or more of the nanoparticles.
The present invention also provides a method for preparing a microneedle patch according to any one of the above embodiments, comprising the steps of:
(1) carrying out ozone treatment on the microneedle patch;
(2) and (2) loading the nano-particles on the microneedle patch treated in the step (1).
Preferably, in the step (1), the ozone treatment concentration is not less than 50g/m3。
Preferably, in the step (1), the ozone treatment time is not less than 5 min.
Preferably, the microneedle patch is a polymer hydrogel formed by covalent crosslinking polymerization of monomers and polymers.
Preferably, in the step (2), the loading concentration of the nanoparticles is not lower than 1nmol/L, and the loading time of the nanoparticles is 5-30 min.
The invention also provides application of the microneedle patch in any scheme or the microneedle patch prepared by the preparation method in any scheme, and the application is applied to detection of microorganisms in food.
Preferably, the microneedle patch enriches microorganisms in food, and combines raman spectroscopy to detect the content of the microorganisms in the food. The micro-needle patch is combined with a Raman spectrum technology, can quickly extract and utilize Raman signals to evaluate the content of microorganisms in food, does not need a pretreatment process, and has simple and convenient operation and high sensitivity.
Compared with the prior art, the invention has the beneficial effects that:
1. the microneedle patch provided by the invention can effectively load nanoparticles with a plasma resonance effect by performing efficient surface plasma treatment on a chemically crosslinked microneedle body, and has the advantages of simplicity and convenience in operation, short extraction time, capability of being stored at normal temperature and the like.
2. The microneedle patch disclosed by the invention is applied to rapidly enriching analytes to be detected in a solid sample, can be used for rapidly extracting and utilizing Raman signals to evaluate the content of microorganisms in food by combining a Raman spectrum technology, does not need a pretreatment process, and is simple and convenient to operate, high in sensitivity and wide in application prospect.
Drawings
Fig. 1 is a flowchart of the preparation of a microneedle patch according to a first embodiment of the present invention;
fig. 2 is a digital photograph of a microneedle patch according to a first embodiment of the present invention;
fig. 3 is a scanning electron microscope image of a microneedle patch according to a first embodiment of the present invention;
FIG. 4 is a schematic diagram of a principle of micro-needle patch extraction of microorganisms according to a first embodiment of the present invention;
FIG. 5 is a surface enhanced Raman spectrum of E.coli extracted by microneedle patches according to one embodiment of the present invention.
Detailed Description
The invention provides a microneedle patch with a plasma enhancement effect, a preparation method and an application thereof. In order to more clearly illustrate the embodiments of the present invention, the technical solutions provided by the present invention will be described in detail with reference to the accompanying drawings, but they should not be construed as limiting the scope of the present invention.
The first embodiment is as follows:
as shown in fig. 1, the method for preparing a microneedle patch with a plasma enhanced effect of the present embodiment includes the following steps:
1. preparation of chemically crosslinked microneedle patch
(1) Preparing a mixed solution of a comonomer, a crosslinking agent and an initiator: mixing ethylene glycol, ethanol and 3mol/L sodium hydroxide solution according to the weight ratio of 1.5: 1: 1.5 volume ratio to 400 u L solution; adding 1mmol of [2- (methacryloyloxy) ethyl ] trimethyl ammonium chloride (TM), 1mmol of 2-carboxyethyl acrylate (CAA), 6mmol of 2-hydroxyethyl methacrylate (HEMA) and 4mmol of triethylene glycol dimethyl acrylic acid methyl ester (TEGDMA) into the solution, and mixing; respectively taking 16 mu L of 40 wt% ammonium persulfate solution and 15 wt% sodium metabisulfite solution, adding the ammonium persulfate solution and the 15 wt% sodium metabisulfite solution into the mixed solution, and fully mixing to obtain a mixed solution;
(2) filling the mixed solution in a microneedle template and carrying out chemical crosslinking: injecting the mixed solution into a microneedle template to fully fill the microneedle template and remove redundant liquid and bubbles on the surface; placing the microneedle template which is filled with the microneedles and is removed with redundant liquid and bubbles on the surface into a 60 ℃ oven for heat treatment, wherein the heat treatment time is 1 h; after complete polymerization, the microneedle patch was carefully removed from the microneedle template with forceps to achieve demolding.
2. Loading nano-particles after demoulding and ozone treatment
(1) Preparation of Nanogold colloid
All glassware used in this experiment was first soaked and cleaned with aqua regia for use.
47.5mL of ultrapure water was weighed outPutting water into a conical flask, placing on a magnetic stirrer at a set rotating speed of 600r/min, and uniformly adding 1.25mL of HAuCl at a concentration of 4g/L4And (3) solution.
The solution in the flask was heated to boiling with stirring at an elevated temperature of 260 ℃. After boiling for 3min, 2mL of 10mg/mL aqueous sodium citrate solution was added.
And stopping heating after the solution is converted from colorless to blue and purple and finally becomes wine red, continuously stirring and cooling to room temperature to obtain the freshly prepared nano gold colloid, and standing at 4 ℃ for later use.
The concentration of the nano-gold is 1.19nmol/L according to the absorbance and the Lambert beer law.
(2) Ozone treatment
Placing the microneedle patch in an ozone generator with ozone concentration of 100g/m3The treatment time is 5 min.
(3) Loaded with nano gold
Immersing the microneedle patch in the nanogold colloid with the needle tip upward, and incubating for 25 min; and finally, gently taking out the microneedle patch by using forceps, and drying the microneedle patch in a drying dish to obtain the microneedle patch with the surface combined with a large amount of nano gold, namely the microneedle patch with the plasma enhancement effect.
The microneedle patch of the embodiment has a surface plasma enhancement effect through surface plasma treatment, and can be applied to the SERS detection technology.
The microneedle patch prepared by the embodiment is applied to rapidly extracting escherichia coli in food, and qualitative detection is carried out through SERS spectrum.
Specifically, the types of the microneedle templates shown in table 1 were selected.
TABLE 1 type of microneedle template selected for use in this example
TABLE 2 sequences of E.coli aptamers selected for use in this example
The sequence of the Escherichia coli aptamer is shown in Table 2, and the Escherichia coli aptamer is dissolved in ultrapure water at 25 ℃ to prepare an aptamer solution with the concentration of 0.25 mu mol/L; incubating the microneedle patch prepared in the first embodiment in the aptamer solution for 25 min; then, the sample was gently removed with tweezers and dried in a drying dish.
Adding nutrient agar powder with the content of 15 wt% into the escherichia coli suspension, and cooling to obtain the culture medium artificially inoculated with the escherichia coli.
Extraction of Escherichia coli: and sticking a microneedle patch on the surface of the culture medium artificially inoculated with the escherichia coli, slightly pressing for 5min by using forceps, and then peeling the microneedle patch for later use.
Fig. 4 is a schematic diagram illustrating the principle of extracting escherichia coli by the microneedle patch loaded with a large amount of nanogold on the surface in this embodiment.
And (3) detection of escherichia coli: and (3) qualitatively detecting the escherichia coli extracted from the microneedle patch by using laser Raman spectroscopy. Collecting 500-1900cm under 785nm excitation light-1SERS signal over range.
The curve is plotted with the Raman intensity as abscissa and the relative intensity as ordinate, as shown in FIG. 5, E.coli at 602, 653, 958, 1450, 1578, 1805cm-1Has Raman vibration peaks, and the peak intensity of the characteristic peaks is obviously enhanced.
The microneedle patch of the embodiment is used as an active substrate for SERS detection, a pretreatment process is not needed, the operation is simple and convenient, the use is easy, the nanogold with the surface plasma resonance effect is loaded on the surface of the needle body, the Raman detection signal is greatly enhanced, the microneedle patch is suitable for the evaluation of the content of microorganisms in food, and the microneedle patch has a strong application prospect and a very high popularization value.
In the above embodiment and its alternatives, the ozone treatment concentration may also be 50g/m3The above is arbitrarily selected.
In the above embodiments and alternatives, the ozone treatment time can also be arbitrarily selected above 5 min.
In the above embodiments and alternatives thereof, the nanoparticle loading concentration can also be arbitrarily selected above 1 nmol/L.
In the above embodiments and alternatives thereof, the nanoparticle loading can be arbitrarily selected within the range of 5-30 min.
In the above embodiments and alternatives thereof, the nanoparticles may also be nanogold, nanosilver, and CuO, Al2O3、MgO、Fe2O3、ZnO、ZrO2One or more of the nano particles are selected according to the actual application requirements.
In view of the numerous embodiments of the present invention, the experimental data of each embodiment is huge and is not suitable for being listed and explained herein one by one, but the contents to be verified and the final conclusions obtained by each embodiment are close. Therefore, the contents of the verification of each embodiment are not described one by one, and only the embodiment one is taken as a representative to illustrate the superiority of the present invention.
The foregoing is merely a preferred embodiment of the invention, and it should be noted that those skilled in the art can make changes in the detailed description without departing from the principles of the invention, and that such changes should be considered as within the scope of the invention.
Claims (10)
1. A micro-needle patch with a plasma enhancement effect is characterized in that the micro-needle patch is used as a carrier, and nanoparticles with a plasma resonance effect are loaded on the surface of the micro-needle patch.
2. A microneedle patch having a plasma-enhanced effect according to claim 1, wherein said microneedle patch has a chemical cross-linking structure.
3. A microneedle patch having a plasma enhanced effect according to claim 1, wherein said nanoparticles are nanogold, nanosilver, CuO, Al2O3、MgO、Fe2O3、ZnO、ZrO2In the nano-particlesOne or more of (a).
4. A method of manufacturing a microneedle patch according to claim 1, 2 or 3, comprising the steps of:
(1) carrying out ozone treatment on the microneedle patch;
(2) and (2) loading the nano-particles on the microneedle patch treated in the step (1).
5. The production method according to claim 4, wherein in the step (1), the ozone treatment concentration is not less than 50g/m3。
6. The method according to claim 4, wherein in the step (1), the ozone treatment time is not less than 5 min.
7. The method of manufacturing according to claim 4, wherein the microneedle patch is a polymer hydrogel formed by covalent crosslinking polymerization of monomers and polymers.
8. The preparation method according to claim 4, wherein in the step (2), the loading concentration of the nanoparticles is not less than 1nmol/L, and the loading time of the nanoparticles is 5-30 min.
9. Use of the microneedle patch according to any one of claims 1 to 3 or the microneedle patch produced by the production method according to any one of claims 4 to 8 for detection of microorganisms in foods.
10. The use of claim 9, wherein the microneedle patch is enriched for microorganisms in food products, in combination with raman spectroscopy, to detect the microbial content of the food products.
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