CN112321870B - Microneedle patch and preparation method and application thereof - Google Patents
Microneedle patch and preparation method and application thereof Download PDFInfo
- Publication number
- CN112321870B CN112321870B CN202011227837.1A CN202011227837A CN112321870B CN 112321870 B CN112321870 B CN 112321870B CN 202011227837 A CN202011227837 A CN 202011227837A CN 112321870 B CN112321870 B CN 112321870B
- Authority
- CN
- China
- Prior art keywords
- microneedle
- microneedle patch
- metal oxide
- needle body
- ultrapure water
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J7/00—Chemical treatment or coating of shaped articles made of macromolecular substances
- C08J7/04—Coating
- C08J7/043—Improving the adhesiveness of the coatings per se, e.g. forming primers
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09D—COATING COMPOSITIONS, e.g. PAINTS, VARNISHES OR LACQUERS; FILLING PASTES; CHEMICAL PAINT OR INK REMOVERS; INKS; CORRECTING FLUIDS; WOODSTAINS; PASTES OR SOLIDS FOR COLOURING OR PRINTING; USE OF MATERIALS THEREFOR
- C09D129/00—Coating compositions based on homopolymers or copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and at least one being terminated by an alcohol, ether, aldehydo, ketonic, acetal, or ketal radical; Coating compositions based on hydrolysed polymers of esters of unsaturated alcohols with saturated carboxylic acids; Coating compositions based on derivatives of such polymers
- C09D129/02—Homopolymers or copolymers of unsaturated alcohols
- C09D129/04—Polyvinyl alcohol; Partially hydrolysed homopolymers or copolymers of esters of unsaturated alcohols with saturated carboxylic acids
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09D—COATING COMPOSITIONS, e.g. PAINTS, VARNISHES OR LACQUERS; FILLING PASTES; CHEMICAL PAINT OR INK REMOVERS; INKS; CORRECTING FLUIDS; WOODSTAINS; PASTES OR SOLIDS FOR COLOURING OR PRINTING; USE OF MATERIALS THEREFOR
- C09D7/00—Features of coating compositions, not provided for in group C09D5/00; Processes for incorporating ingredients in coating compositions
- C09D7/40—Additives
- C09D7/60—Additives non-macromolecular
- C09D7/61—Additives non-macromolecular inorganic
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2327/00—Characterised by the use of homopolymers or copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and at least one being terminated by a halogen; Derivatives of such polymers
- C08J2327/02—Characterised by the use of homopolymers or copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and at least one being terminated by a halogen; Derivatives of such polymers not modified by chemical after-treatment
- C08J2327/12—Characterised by the use of homopolymers or copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and at least one being terminated by a halogen; Derivatives of such polymers not modified by chemical after-treatment containing fluorine atoms
- C08J2327/18—Homopolymers or copolymers of tetrafluoroethylene
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2429/00—Characterised by the use of homopolymers or copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and at least one being terminated by an alcohol, ether, aldehydo, ketonic, acetal, or ketal radical; Hydrolysed polymers of esters of unsaturated alcohols with saturated carboxylic acids; Derivatives of such polymer
- C08J2429/02—Homopolymers or copolymers of unsaturated alcohols
- C08J2429/04—Polyvinyl alcohol; Partially hydrolysed homopolymers or copolymers of esters of unsaturated alcohols with saturated carboxylic acids
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08K—Use of inorganic or non-macromolecular organic substances as compounding ingredients
- C08K3/00—Use of inorganic substances as compounding ingredients
- C08K3/18—Oxygen-containing compounds, e.g. metal carbonyls
- C08K3/20—Oxides; Hydroxides
- C08K3/22—Oxides; Hydroxides of metals
- C08K2003/2237—Oxides; Hydroxides of metals of titanium
- C08K2003/2241—Titanium dioxide
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08K—Use of inorganic or non-macromolecular organic substances as compounding ingredients
- C08K3/00—Use of inorganic substances as compounding ingredients
- C08K3/18—Oxygen-containing compounds, e.g. metal carbonyls
- C08K3/20—Oxides; Hydroxides
- C08K3/22—Oxides; Hydroxides of metals
- C08K2003/2293—Oxides; Hydroxides of metals of nickel
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08K—Use of inorganic or non-macromolecular organic substances as compounding ingredients
- C08K3/00—Use of inorganic substances as compounding ingredients
- C08K3/18—Oxygen-containing compounds, e.g. metal carbonyls
- C08K3/20—Oxides; Hydroxides
- C08K3/22—Oxides; Hydroxides of metals
- C08K2003/2296—Oxides; Hydroxides of metals of zinc
Abstract
The invention belongs to the technical field of food allergen detection, and particularly relates to a preparation method of a microneedle patch, which comprises the following steps: dispersing metal oxide in MES buffer solution to prepare metal oxide solution; adding polyvinyl alcohol into ultrapure water, adding the ultrapure water into a metal oxide solution, and stirring to prepare a needle body preparation liquid; coating the microneedle template with the needle body manufacturing liquid so as to fill the microneedle template with the needle body manufacturing liquid; carrying out positive pressure treatment, vacuum treatment and constant temperature drying on the microneedle template, and then demoulding to obtain a microneedle patch; the microneedle patch disclosed by the invention is applied to rapid extraction of DNA, can shorten the extraction time, can be directly used for semi-quantitatively detecting the allergen in deep-processed food without purification of the extracted DNA, has the advantages of being minimally invasive and short in extraction time, is simple and convenient to operate, does not need purification, and is convenient for subsequent operation.
Description
Technical Field
The invention belongs to the technical field of food allergen detection, and particularly relates to a microneedle patch as well as a preparation method and application thereof.
Background
Food allergies are a common type of health problem in human daily life. To date, the detection of food allergens has been largely divided into two categories, one in which the allergen is detected directly against the allergenic protein and the other in which a DNA fragment is indirectly indicative of the presence of the allergenic component. Although DNA is readily degraded, in a food matrix, DNA remains stable under heat and pressure treatment conditions for a longer period of time than protein, and target DNA degrades at a much slower rate than protein, so DNA characteristic of the residual component allergen is better indicative of the presence of the allergen.
The deep processed food contains various raw material components, and also contains food additives such as salt, sugar, pigment, etc. due to complex food components. In addition, the sensitizing protein in the raw materials can be damaged to different degrees by the processes of frying, boiling and the like in the processing process, so that the difficulty of the method for directly detecting the allergen through the sensitizing protein is increased. At present, the most common methods for specifically detecting allergen DNA in food are PCR and qRT-PCR technologies, but before the detection method is used, various lysates are generally required to extract DNA, the time is consumed for about 4-5 h, and the application of the method is limited. Although commercial kits for extracting DNA from deep-processed food are available in the market at present, the whole process still needs 2-3 hours, so that the research of a rapid pretreatment method becomes an important direction for expanding the application range of detection technology.
Microneedle patches, i.e., needle-like projections having micron-scale dimensions, are typically assembled in an array onto a supporting substrate or patch. In view of the current domestic and foreign literature reports, the microneedle patch is successfully applied to the aspects of vaccination, diabetes treatment, skin disease treatment and the like, and in recent years, researches show that the microneedle patch has good extraction capacity in the aspect of absorbing interstitial fluid of the skin by using the swelling capacity of the microneedle patch without an external instrument, so that the microneedle patch has certain application potential in the rapid separation and extraction of food characteristic DNA.
At present, materials such as silicon materials, metal materials, high polymer materials and the like have been proved to be capable of being used for preparing microneedle patches, wherein the high polymer materials are widely used for preparing the microneedle patches due to the advantages of wide raw material sources, low price, relatively simple preparation method, suitability for industrial production and the like. Polyvinyl alcohol (PVA) is used as a high-molecular material with strong water absorption, and has good biocompatibility, chemical resistance and mechanical strength. PVA gel and derivatives thereof are highly expansive materials, and can rapidly absorb 10-30% of water by self weight within a few minutes, so that polyvinyl alcohol becomes a hotspot for research of microneedle materials.
Since there is an unavoidable problem in extracting DNA with microneedle patches, the concentration of target nucleic acid is generally much lower than that of coexisting proteins. According to the current domestic and foreign literature reports, many nano materials can strongly adsorb DNA, including various metal oxides, graphene oxide, polymeric ionic liquid, ionic resin, chitosan, cellulose and the like. Among the above materials, metal oxides can interact well with the phosphate skeleton of DNA, and thus, the use of metal oxides in the fabrication of microneedle patches is expected to improve the ability of microneedle patches to extract DNA.
Through the current description of the allergen detection technology and the introduction of the preparation material of the microneedle patch and the advantages of polyvinyl alcohol and metal oxide, for this reason, it is necessary to develop a microneedle patch for rapidly extracting the characteristic DNA of the allergen in deep-processed food in a water-absorbing swelling manner, so as to overcome the defects in the practical application of rapid food allergen detection.
Disclosure of Invention
Based on the above-mentioned disadvantages and drawbacks of the prior art, an object of the present invention is to solve at least one or more of the above-mentioned problems of the prior art, in other words, to provide a method for manufacturing a microneedle patch that satisfies one or more of the above-mentioned needs.
It is another object of the present invention to provide a microneedle patch.
The invention also aims to provide an application scheme of the microneedle patch.
In order to achieve the purpose, the invention adopts the following technical scheme:
a method for preparing a microneedle patch, comprising the steps of:
(1) Dispersing a metal oxide in MES (2- (N-morpholine) ethanesulfonic acid) buffer at 25 ℃ to prepare a metal oxide solution;
(2) Adding polyvinyl alcohol into ultrapure water, adding the polyvinyl alcohol into a metal oxide solution, stirring by using a magnetic stirrer until PVA is completely dissolved in the water, and standing for several hours to obtain a clear and transparent needle body preparation liquid with slight fluidity;
(3) Putting a Polydimethylsiloxane (PDMS) microneedle template into a beaker filled with ultrapure water, boiling and cleaning, then putting the PDMS microneedle template into an ethanol solution, and cleaning with an ultrasonic cleaning machine; coating the obtained needle body manufacturing liquid in a clean and dry PDMS microneedle template, and scraping redundant manufacturing liquid and bubbles on the surface of the microneedle mould;
(4) And (3) carrying out positive pressure treatment, vacuum treatment and constant temperature drying on the microneedle template, and then carefully peeling the microneedle patch from the microneedle template by using tweezers to obtain the microneedle patch.
Preferably, the needle body preparation solution has a mass of 12 to 16wt%, a volume of the metal oxide solution of 10 to 15vol%, and a concentration of the metal oxide solution of 0.2mg/mL.
Preferably, the metal oxide is one or more of nickel oxide, zinc oxide and titanium dioxide.
Preferably, the temperature of the ultrapure water is 90-100 ℃.
Preferably, the stirring speed is not lower than 1000rpm, and the stirring temperature is not lower than 90 ℃.
Preferably, the constant-temperature drying temperature is 40-60 ℃, and the drying time is not less than 8h.
Preferably, the pressure of the vacuum treatment is not less than 0.02MPa, the temperature is 30-60 ℃, and the vacuum time is not less than 30min.
Preferably, the step (3) specifically comprises: and (3) cleaning the PDMS microneedle template by ultrapure water and ultrasonic cleaning, coating the needle body manufacturing liquid in the dried PDMS microneedle template, and scraping redundant microneedle manufacturing liquid and bubbles.
The invention also provides the microneedle patch prepared by the preparation method according to any one of the above schemes.
The invention also provides the microneedle patch prepared by the preparation method in any scheme or the application of the microneedle patch in the above scheme, and the microneedle patch is applied to extraction of deep-processing food allergen DNA.
Compared with the prior art, the invention has the beneficial effects that:
1. the raw material polyvinyl alcohol is non-toxic and harmless, can be biologically degraded, is not easy to cause pollution, and has strong water absorption, good biocompatibility, chemical resistance and mechanical strength.
2. The microneedle patch is prepared by taking metal oxide and polyvinyl alcohol as raw materials, drying and demolding by a special positive pressure and vacuum process, has the advantages of minimal invasion and short extraction time, is simple and convenient to operate, does not need purification and is convenient for subsequent operation.
3. The microneedle patch disclosed by the invention is applied to rapid extraction of DNA, the pretreatment time can be obviously shortened to 2min from 2-3 h of a commercialized kit method, and the extracted DNA can be directly used for semi-quantitatively detecting the allergen in deep-processed food without purification, is suitable for rapid field detection of the food allergen, and has a wide 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 schematic diagram of the principle of DNA extraction by microneedle patches according to the second embodiment of the present invention;
FIG. 4 is a PCR electrophoretogram of a second allergen detection kit according to the present invention;
in the figure: 1 purpose detection food, 2 free DNA, 3 cross-linked PVA matrix and 4 nanometer metal oxide.
Detailed Description
In order to more clearly illustrate the embodiments of the present invention, the following description will explain the embodiments of the present invention with reference to the accompanying drawings. It is obvious that the drawings in the following description are only some examples of the invention, and that for a person skilled in the art, other drawings and embodiments can be derived from them without inventive effort.
The first embodiment is as follows:
as shown in fig. 1 and fig. 2, this embodiment provides a method for preparing a microneedle patch, including the following steps:
(1) Nickel oxide or zinc oxide was dispersed in MES (2- (N-morpholino) ethanesulfonic acid) buffer at 25 ℃ to prepare a metal oxide solution with a concentration of 0.2mg/mL.
(2) Adding 15wt% of polyvinyl alcohol (PVA) into ultrapure water at 95 ℃, adding 10vol% of nickel oxide or zinc oxide solution, stirring by using a magnetic stirrer at 95 ℃ and 1000rpm until the PVA is completely dissolved in the water, and standing for several hours to obtain clear and transparent fluid needle body preparation liquid.
(3) Putting a Polydimethylsiloxane (PDMS) microneedle template into a beaker filled with ultrapure water, boiling and cleaning for 5min, then putting the PDMS microneedle template into a 75% ethanol solution, cleaning for 5min by using an ultrasonic cleaner, coating the obtained needle body manufacturing liquid in the clean and dry PDMS microneedle template, and scraping off the excess microneedle manufacturing liquid and bubbles on the surface of the PDMS microneedle template.
(4) Putting the PDMS microneedle template filled with the microneedle preparation liquid into a self-made positive pressure needle cylinder device for pressing treatment so as to remove small bubbles in the microneedle preparation liquid, then putting the microneedle preparation liquid into a vacuum drying oven for vacuum treatment under the vacuum condition of 0.02MPa, the vacuum drying temperature of 30 ℃ and the vacuum time of 30min so as to remove the residual bubbles and completely fill a microneedle cavity with the microneedle preparation liquid, and finally putting the microneedle template subjected to vacuum treatment into a blast drying oven for drying at constant temperature of 40 ℃ for 8h. And (3) after the microneedle patch is completely dried, filling the microneedle template with the microneedle patch by using tweezers, and peeling off the microneedle template to obtain the microneedle patch with swelling capacity.
The raw material polyvinyl alcohol of the microneedle patch of the embodiment is nontoxic and harmless, is biodegradable, is not easy to cause pollution, and has strong water absorption, good biocompatibility, chemical resistance and mechanical strength.
Example two:
the embodiment provides an application of a microneedle patch, which is used for quickly extracting allergen DNA from self-made milk pudding (agar base) and performing qualitative detection through PCR.
In consideration of the hardness and shape of the milk pudding, the model of the microneedle mold plate shown in table 1 was selected for this example, and the mass fraction of the needle body-producing fluid was selected to be 13wt%.
TABLE 1 type of microneedle template selected for use in this example
The preparation process of the milk pudding comprises the following steps: adding agar powder with content of 0.8wt% into fresh milk, boiling, and cooling to obtain milk pudding.
The microneedle patch was attached to the surface of a milk pudding, gently pressed with a hand to transfer the piercing force to the patch, and the swelling uptake time was 1min. Stripping the microneedle patch and washing the microneedle patch for multiple times by 50 mu L of TE buffer to obtain a solution which is a DNA template used for subsequent PCR, and performing PCR amplification by using a specific primer. The allergen in the milk pudding mainly comprises alpha-lactalbumin, beta-lactoglobulin and beta-casein in milk, wherein specific primers used for PCR are shown in Table 2.
TABLE 2 primer sequences for DNA characteristic of each allergen
As shown in fig. 3, the principle process of extracting DNA by microneedle patch added with nano metal oxide in this embodiment is shown. The amplification reaction was performed in a volume of 50. Mu.L, and the DNA template was replaced with ultrapure water as a negative control. The PCR reaction system consists of 2. Mu.l DNA template, 2. Mu.l upstream primer, 2. Mu.l downstream primer, 25. Mu.l
Max Buffer、1μl dNTP Mix、17μl ddH 2 O and 1. Mu.l
Max Super-Fidelity DNA Polymerase. Centrifugally and uniformly mixed and placed on a PCR instrument for carrying outAutomated amplification reactions.
The PCR circulation conditions suitable for the specific primers of different allergens are different, so the alpha-lactalbumin circulation conditions are 94 ℃ for initial template denaturation and 3min; denaturation at 94 ℃ for 30s; annealing at 55 deg.C for 30s, stretching at 72 deg.C for 30s, and stretching at 72 deg.C for 10min after 35 cycles. The circulation condition of the beta-lactoglobulin is that the initial template is denatured at 95 ℃ for 3min; denaturation at 95 ℃ for 30s; annealing at 55 deg.C for 30s, stretching at 72 deg.C for 30s, and stretching at 72 deg.C for 10min after 30 cycles. The beta-casein circulating condition is that the initial template is denatured at 95 ℃ for 5min; denaturation at 95 ℃ for 30s; annealing at 64 deg.C for 30s, stretching at 72 deg.C for 30s, and stretching at 72 deg.C for 5min after 43 cycles.
5 mul of PCR amplification product is spotted in 1.5% agarose gel for detection, the voltage is 160V, 169min, then imaging is carried out in a gel imaging system, and whether the PCR amplification product has the expected DNA specific fragment or not is observed.
The results of PCR electrophoresis are shown in FIG. 4, which shows that the microneedle patch can extract DNA characteristic to food, and that the capability of both extracting DNA characteristic to milk pudding allergen is similar to that of the kit.
The method for detecting the microneedle patch is simple and convenient to operate, easy to use, capable of semi-quantitatively detecting the allergen DNA in food and high in popularization value.
The microneedle patch provided by the invention can be used as a method for rapidly extracting DNA, the pretreatment time can be remarkably shortened to 2min from 2-3 h of a commercial kit method, and the extracted DNA can be directly used for semi-quantitatively detecting the allergen in deep-processed food without purification, is suitable for rapid field detection of the food allergen, and has a wide application prospect.
In the above embodiment and the alternative, the needle body manufacturing liquid may be arbitrarily selected in a range of 12 to 16 wt%; the volume of the metal oxide solution may be arbitrarily selected within the range of 10 to 15 vol%.
In the above embodiment and its alternatives, the stirring speed can also be arbitrarily chosen above 1000 rpm; the stirring temperature can be arbitrarily selected above 90 ℃; the temperature for constant temperature drying can be arbitrarily selected within the range of 40-60 ℃; the constant-temperature drying time can be selected at will for more than 8 hours; the pressure intensity of vacuum drying can be arbitrarily selected above 0.02 MPa; the vacuum temperature can be arbitrarily selected within the range of 30-60 ℃; the vacuum time can be arbitrarily selected above 30min.
In the above embodiment and its alternatives, the temperature of the ultrapure water can be arbitrarily selected within the range of 90 to 100 ℃; the boiling and cleaning time in the ultrapure water can be randomly selected within the range of 5-10 min; the ultrasonic cleaning time can be randomly selected within 5-10 min.
The foregoing has outlined, rather broadly, the preferred embodiment and principles of the present invention in order that those skilled in the art may better understand the detailed description of the invention without departing from its broader aspects.
Claims (8)
1. A method for preparing a microneedle patch is characterized by comprising the following steps:
(1) Dispersing metal oxide in MES buffer solution to prepare metal oxide solution;
the metal oxide is one or more of nickel oxide, zinc oxide and titanium dioxide;
(2) Adding polyvinyl alcohol into ultrapure water, adding the polyvinyl alcohol into a metal oxide solution, and stirring to prepare a needle body preparation liquid;
the mass of the polyvinyl alcohol in the needle body manufacturing liquid is 12-16 wt% of the mass of the ultrapure water, and the volume of the metal oxide solution is 10-15 vol% of the volume of the ultrapure water;
(3) Coating the microneedle mould plate with the needle body manufacturing liquid so as to fill the microneedle mould plate with the needle body manufacturing liquid;
(4) And (3) carrying out positive pressure treatment, vacuum treatment and constant temperature drying on the microneedle template, and then demoulding to obtain the microneedle patch.
2. A method for preparing a microneedle patch according to claim 1, wherein the ultrapure water is heated at 90 to 100 ℃.
3. A method for preparing a microneedle patch according to claim 1, wherein the stirring speed is not less than 1000rpm and the stirring temperature is not less than 90 ℃.
4. The method for preparing a microneedle patch according to claim 1, wherein the drying temperature at constant temperature is 40-60 ℃ and the drying time at constant temperature is not less than 8 hours.
5. A method for preparing a microneedle patch according to claim 1, wherein the pressure of the vacuum treatment is not less than 0.02MPa, the temperature is 30-60 ℃, and the vacuum time is not less than 30min.
6. A method for preparing a microneedle patch according to claim 1, wherein the step (3) specifically comprises: and (3) cleaning the PDMS microneedle template by ultrapure water and ultrasonic cleaning, coating the needle body manufacturing liquid in the dried PDMS microneedle template, and scraping redundant microneedle manufacturing liquid and bubbles.
7. A microneedle patch characterized by being produced by the production method according to any one of claims 1 to 6.
8. Use of a microneedle patch according to claim 7 for extracting DNA from deep processed food allergens.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011227837.1A CN112321870B (en) | 2020-11-06 | 2020-11-06 | Microneedle patch and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011227837.1A CN112321870B (en) | 2020-11-06 | 2020-11-06 | Microneedle patch and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN112321870A CN112321870A (en) | 2021-02-05 |
| CN112321870B true CN112321870B (en) | 2022-11-18 |
Family
ID=74316675
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011227837.1A Active CN112321870B (en) | 2020-11-06 | 2020-11-06 | Microneedle patch and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112321870B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113358625A (en) * | 2021-05-14 | 2021-09-07 | 浙江工商大学 | Microneedle patch with plasma enhancement effect and preparation method and application thereof |
| CN113995444A (en) * | 2021-11-03 | 2022-02-01 | 深圳大学 | Hydrogel microneedle patch and preparation method and application thereof |
| CN116350583A (en) * | 2023-03-30 | 2023-06-30 | 沈阳药科大学 | Zinc oxide nano particle/ionic liquid dispersion with synergistic antibacterial effect and preparation and application thereof |
Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001034844A1 (en) * | 1999-11-10 | 2001-05-17 | Ligochem, Inc. | Method for isolating dna from a proteinaceous medium and kit for performing method |
| CN101175853A (en) * | 2005-03-16 | 2008-05-07 | Dna吉诺特克股份有限公司 | Compositions and method for storage of nucleic acid from bodily fluids |
| CN101613696A (en) * | 2009-08-05 | 2009-12-30 | 公安部物证鉴定中心 | Extract the reagent of purify DNA |
| CN101613697A (en) * | 2009-08-05 | 2009-12-30 | 公安部物证鉴定中心 | A kind of method of extracting purify DNA |
| CN101627065A (en) * | 2006-12-21 | 2010-01-13 | 茵维特罗根戴纳股份公司 | Particle and its application in the method for the method of isolating nucleic acid or isolating phosphoproteins |
| CN102115739A (en) * | 2009-12-31 | 2011-07-06 | 财团法人工业技术研究院 | Method, reagent and kit for separating nucleic acids |
| CN102220310A (en) * | 2009-08-05 | 2011-10-19 | 公安部物证鉴定中心 | Method for extracting and purifying spittle DNA |
| CN103789297A (en) * | 2012-10-26 | 2014-05-14 | 上海医脉赛科技有限公司 | Nucleic acid rapid-purifying method and kit |
| CN104066475A (en) * | 2012-02-17 | 2014-09-24 | 考司美德制药株式会社 | Microneedle of short-time dissolution type |
| CN109364366A (en) * | 2018-09-21 | 2019-02-22 | 华中科技大学 | Template prepares the method and its application of porous polymer micropin |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040063100A1 (en) * | 2002-09-30 | 2004-04-01 | Wang Chung Lin | Nanoneedle chips and the production thereof |
-
2020
- 2020-11-06 CN CN202011227837.1A patent/CN112321870B/en active Active
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001034844A1 (en) * | 1999-11-10 | 2001-05-17 | Ligochem, Inc. | Method for isolating dna from a proteinaceous medium and kit for performing method |
| CN101175853A (en) * | 2005-03-16 | 2008-05-07 | Dna吉诺特克股份有限公司 | Compositions and method for storage of nucleic acid from bodily fluids |
| CN101627065A (en) * | 2006-12-21 | 2010-01-13 | 茵维特罗根戴纳股份公司 | Particle and its application in the method for the method of isolating nucleic acid or isolating phosphoproteins |
| CN101613696A (en) * | 2009-08-05 | 2009-12-30 | 公安部物证鉴定中心 | Extract the reagent of purify DNA |
| CN101613697A (en) * | 2009-08-05 | 2009-12-30 | 公安部物证鉴定中心 | A kind of method of extracting purify DNA |
| CN102220310A (en) * | 2009-08-05 | 2011-10-19 | 公安部物证鉴定中心 | Method for extracting and purifying spittle DNA |
| CN102115739A (en) * | 2009-12-31 | 2011-07-06 | 财团法人工业技术研究院 | Method, reagent and kit for separating nucleic acids |
| CN104066475A (en) * | 2012-02-17 | 2014-09-24 | 考司美德制药株式会社 | Microneedle of short-time dissolution type |
| CN103789297A (en) * | 2012-10-26 | 2014-05-14 | 上海医脉赛科技有限公司 | Nucleic acid rapid-purifying method and kit |
| CN109364366A (en) * | 2018-09-21 | 2019-02-22 | 华中科技大学 | Template prepares the method and its application of porous polymer micropin |
Non-Patent Citations (2)
| Title |
|---|
| DNA Extraction from Plant Leaves Using a Microneedle Patch;Rajesh Paul等;《Current Protocols in Plant Biology》;20200219;第5卷;第1-12页 * |
| Extraction of Plant DNA by Microneedle Patch for Rapid Detection of Plant Diseases;Rajesh Paul等;《ACS Nano》;20190609;第13卷;第6540-6549页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN112321870A (en) | 2021-02-05 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN112321870B (en) | Microneedle patch and preparation method and application thereof | |
| Liu et al. | Effects of in vitro digestion and fecal fermentation on the stability and metabolic behavior of polysaccharides from Craterellus cornucopioides | |
| CN101704964A (en) | Medical security level rubber product with sericin modified surface and preparation method thereof | |
| FR2597481A1 (en) | PROCESS FOR OBTAINING CHEMICALLY DEFINED AND REPRODUCIBLE POLYDESOXYRIBONUCLEOTIDES | |
| CN111285925B (en) | Separation and purification method of lactobacillus paracasei ZFM54 bacteriocin | |
| CN111020001A (en) | Novel saliva preserving fluid and preparation method and application thereof | |
| CN107141517A (en) | A kind of method for preparing degradable plastic film using discarded shrimp shell | |
| CN105039480A (en) | Method for purifying collagen derived from pelodiscus sinensis | |
| CN109965173B (en) | Preparation method and application of phycoerythrin, polysaccharide and dietary fiber extracted from laver in water powder | |
| CN110859301A (en) | Preparation method of mesona chinensis benth polysaccharide-soybean protein isolate composite gel | |
| CN113564069B (en) | Bifidobacterium longum, extracellular polysaccharide of bifidobacterium longum, and extraction method and application thereof | |
| JPH03163102A (en) | Preparation of thermally coagulatable beta-1,3-glucan | |
| CN102676568A (en) | Method for producing recombinant dermatophagoides farinae allergen Der f1 and Der f2 fusion protein | |
| CN110082486B (en) | Method for tracing mutton sheep producing area by using stable isotope of ossein | |
| CN110903413B (en) | Jerusalem artichoke pulp inulin for improving functional dyspepsia and preparation method and application thereof | |
| CN107319455B (en) | Wall-broken pollen Pini and its preparation method | |
| CN113155987B (en) | Microneedle patch and preparation method and application thereof | |
| CN110606880B (en) | Combined preparation process of earthworm elastin and lumbrokinase | |
| JP2018174905A (en) | Method for producing lps-containing rice bran extract, and health food using rice bran extract | |
| KR101478615B1 (en) | Method for manufacturing of traditional adhesives using enzyme | |
| CN112903386A (en) | Microneedle patch capable of rapidly extracting phosphoprotein and preparation method and application thereof | |
| CN108531536A (en) | The preparation method and applications of Shelled Turtle Trionyx Sinensis collagen peptide | |
| CN1225370A (en) | Method for radio-grafting medical high molecular aquagel membrane | |
| CN113768081B (en) | Method for removing nitrite in bird's nest | |
| CN109180778A (en) | A method of it improving natural feather polypeptide nano particle solution and rate is made |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |