CN103316615A - Preparation and detection method of magnetic microspheres with visible glucose detection function - Google Patents
Preparation and detection method of magnetic microspheres with visible glucose detection function Download PDFInfo
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- CN103316615A CN103316615A CN2013102372536A CN201310237253A CN103316615A CN 103316615 A CN103316615 A CN 103316615A CN 2013102372536 A CN2013102372536 A CN 2013102372536A CN 201310237253 A CN201310237253 A CN 201310237253A CN 103316615 A CN103316615 A CN 103316615A
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Abstract
The invention discloses a preparation method of magnetic microspheres with a visible glucose detection function. The preparation method comprises the following steps: by utilizing a photonic crystal microsphere as a template, preparing magnetic photonic crystal hydrogel microspheres with reverse opal structures through a template copy method of hydrogel. A detection method comprises the steps that: a glucose molecule identification unit is combined with the magnetic photonic crystal hydrogel microspheres, wherein the existence or number of the glucose molecules is expressed by the displacement or color of the reflection peak of the photonic crystal microsphere; a magnet is used for collecting the magnetic microspheres according to the property that the magnetic microspheres have magnetism to improve the naked eye observation accuracy of the color of the magnetic microspheres. The visible glucose detection method based on magnetic microspheres with visible glucose detection function has the advantages of high sensitivity, strong specificity, simple operation, low cost, accurate observation, and the like.
Description
Technical field
The present invention relates to biomedicine, analyzing and testing field, particularly relate to a kind of glucose visible detection method.
Background technology
Over nearly 10 years, the incidence of disease of China's diabetes rises year by year, has become to be only second to tumour, the cardiovascular afterwards lifelong participation disease of the 3rd large harm health of people.The investigation demonstration, China's diabetes prevalence is up to 9.7%, and that is to say among per 10 adults approximately has 1 diabetic.In following several years, China's diabetes prevalence also can continue to rise.Expert's expectation, by 2015, the diabetic of China will be above 6,000 ten thousand.Diabetes have caused serious financial burden, and data shows, account for population approximately 5% diabetic consumed nearly 10% national health operation expenses, increasing rapidly of diabetes medical expense caused serious social concern.
Diabetes are to prevent medicable disease.The key that reduces distribution of diabetes and disability rate is not only in the treatment of finding early diabetes and the science of carrying out, also is the key that reduces society and diabetic's financial burden.Yet, present stage clinically diagnosing diabetes depend on complex instrument, program is loaded down with trivial details, it is ill that disappearance that adding the custom that has regular physical checkups causes absolutely large number diabetic not find early, incurs loss through delay early treatment.Seek a kind of method that can convenient and swift real-time detection concentration of glucose very urgent.
Summary of the invention
The technical problem that the present invention mainly solves provides a kind of preparation method of the magnetic microsphere with the visual measuring ability of glucose and uses described magnetic microsphere with the visual measuring ability of glucose to carry out the glucose visible detection method, conveniently detect the concentration of glucose in the body fluid, remedied and detected clinically the shortcoming that glucose needs complex instrument at present.
For solving the problems of the technologies described above, the technical scheme that the present invention adopts is:
A kind of preparation method with magnetic microsphere of the visual measuring ability of glucose is provided, and described magnetic microsphere prepares by following steps:
(1) preparation process of magnetic photonic crystal hydrogel microsphere:
Photon crystal micro-ball is added before the hydrogel contain magnetic nano-particle in the aggressiveness solution, add again initator, aggressiveness solution is cured before curing reaction is to hydrogel, obtain the magnetic photonic crystal hydrogel microsphere, the described magnetic photonic crystal hydrogel microsphere that obtains is peeled off, place solvent to remove the photon crystal micro-ball masterplate after the washing, take out after two hours with the pure water washing, then be kept in the pure water;
(2) have the preparation process of the magnetic microsphere of the visual measuring ability of glucose:
Method by the chemical bond coupling, glucose molecule recognition unit phenyl boric acid is coupled on the magnetic photonic crystal hydrogel microsphere that makes in the step (1), make the magnetic microsphere with the visual measuring ability of glucose, the described magnetic microsphere with the visual measuring ability of glucose for preparing with after the buffer solution washing, is measured its reflectance spectrum.
In a preferred embodiments, photon crystal micro-ball described in the preparation method of described magnetic microsphere with the visual measuring ability of glucose is assembled by Nano particles of silicon dioxide and poly-methyl acid methyl esters nano particle or polystyrene nanoparticle.
In a preferred embodiments, described in the preparation method of described magnetic microsphere with the visual measuring ability of glucose before the hydrogel aggressiveness be any one or multiple mixing in acrylamide, methyl methacrylate, poly hydroxy ethyl acrylate, acetylbutyrylcellulose, siloxanes methacrylate, fluorine silicon methacrylate, perfluor ether, N-vinylpyrrolidone, polyvinyl alcohol, GMA or the GDMA.
In a preferred embodiments, the magnetic photonic crystal hydrogel microsphere has counter opal structure described in the preparation method of described magnetic microsphere with the visual measuring ability of glucose.
In a preferred embodiments, the method for curing reaction described in the preparation method of described magnetic microsphere with the visual measuring ability of glucose is specially the UV-irradiation polymerization or uses baking oven to carry out heat cure.
In a preferred embodiments, chemical bond coupling method described in the preparation method of described magnetic microsphere with the visual measuring ability of glucose comprises amino with amino reaction, amino and the reaction of carboxyl or the reaction of carboxyl and hydroxyl.
Another object of the present invention provides the method for the visual detection of a kind of glucose:
The magnetic microsphere that will have a visual measuring ability of glucose mixes with glucose solution in cushioning liquid and carries out the specific binding reaction, react complete after, utilize magnet with free magnetic microsphere polymerization, relatively react the color of front and back magnetic microsphere or by measuring the reflectance spectrum of magnetic microsphere, determine the content of glucose.
In a preferred embodiments, describedly use the glucose solution described in the method that the magnetic microsphere with the visual measuring ability of glucose carries out the visual detection of glucose to be tear, urine, saliva or sweat.
The invention has the beneficial effects as follows:
The magnetic microsphere with the visual measuring ability of glucose by the present invention preparation detects concentration of glucose and compares with the existing method that detects clinically concentration of glucose and have the following advantages:
(1) the present invention carries out the visual detection of glucose take the magnetic photonic crystal hydrogel microsphere as carrier, by visually observing the concentration of glucose that detects in tear, urine or the saliva, need not to extract the symptom that blood can find effectively that concentration of glucose rises in diabetic's Early insulin secretion liquid, convenient and swift and have real-time, greatly improved the possibility of diabetes early detections.
(2) the present invention combines the glucose molecule recognition unit with the magnetic photonic crystal hydrogel microsphere, the Chemical Physics environmental evolution causes the variation of the photon band gap of magnetic microsphere to detect after utilizing the glucose molecule recognition unit in conjunction with glucose molecule, the variation of described photon band gap can bring the variation of described magnetic microsphere color, focusing energy through magnetic microsphere is differentiated by human eye easily, so that testing tool is simplified greatly, testing cost reduces greatly.
(3) magnetic microsphere with the visual measuring ability of glucose provided by the invention is simplified the detection method of concentration of glucose, and eliminated and extracted the pain that blood brings to human body, and without any side effects to human body.
Description of drawings
Fig. 1 is the schematic diagram that uses magnetic microsphere with the visual measuring ability of glucose to carry out the visual detection of glucose among the present invention;
The mark of each parts is as follows in the accompanying drawing: 1, have the magnetic microsphere of the visual measuring ability of glucose, 2, glucose solution, 3, magnet, 4, hydrogel, 5, magnetic nano-particle, 6, glucose molecule, 7, the glucose molecule recognition unit.
The specific embodiment
Below in conjunction with specific embodiment such scheme is described further.Should be understood that these embodiment are not limited to limit the scope of the invention for explanation the present invention.The implementation condition that adopts among the embodiment can be done further adjustment according to the condition of concrete detection, and not marked implementation condition is generally the condition in the normal experiment.Any those skilled in the art in the situation that do not break away from the spirit and scope of the present invention, should make various modifications and change, so protection scope of the present invention should be considered as appended claims limited range.
Fig. 1 is the schematic diagram that uses magnetic microsphere with the visual measuring ability of glucose to carry out the visual detection of glucose among the present invention: contain magnetic nano-particle 5 and hydrogel 4 in the magnetic photonic crystal hydrogel microsphere for preparing, method by the chemical bond coupling is coupled at glucose molecule recognition unit 7 phenyl boric acids in the described magnetic photonic crystal hydrogel microsphere, make the magnetic microsphere 1 with the visual measuring ability of glucose, when carrying out the glucose solution detection, the magnetic microsphere 1 that will have a visual measuring ability of glucose mixes with glucose solution 2 in cushioning liquid and carries out the specific binding reaction, react complete after, utilize magnet 3 with free magnetic microsphere polymerization, relatively react the color of front and back magnetic microsphere or by measuring the reflectance spectrum of magnetic microsphere, determine the content of glucose.
Embodiment 1Use the glucose content in the described magnetic microsphere detection tear:
(1) preparation photon crystal micro-ball
The Nano particles of silicon dioxide of monodispersed 280 nm is carried out centrifugal purification, the mass percent of regulating the colloidal nanoparticles in the nano-particle solution is 20%, after the ultrasonic dispersion as the water in micro-fluidic, the foreign minister adopts the hexadecane that contains surfactant, collect the drop that produces, after it is heating and curing, puts into Muffle furnace and carry out 800 ℃ of calcinings, to improve the stability of photon crystal micro-ball.
(2) preparation magnetic photonic crystal hydrogel microsphere
The photon crystal micro-ball of magnetic photonic crystal hydrogel microsphere preparation in the step (1) is as masterplate, and the method by stencil duplicating obtains.Photon crystal micro-ball is added the Fe that contains light trigger 1173 (0.1%, mass volume ratio) and monodispersed 100nm
3O
4(29:1 in the acrylamide of nano particle and the mixed solution of methylene diacrylamide, mol ratio), after fully soaking, behind high voltage mercury lamp radiation 15min, get the magnetic photonic crystal hydrogel microsphere, resulting magnetic photonic crystal hydrogel microsphere is peeled off, washing is placed on 2h in the solvent, obtains the magnetic photonic crystal hydrogel microsphere of counter opal structure.
(3) modification of glucose molecule recognition unit
The magnetic photonic crystal hydrogel microsphere that step (2) is prepared is at the NaOH(10% of 0.1M N, N, N', the N'-tetramethylethylenediamine) in the solution reduction 1h after, wash with 2-(N-morpholine) ethyl sulfonic acid (MES) cushioning liquid, in the MES buffer solution, add 1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimides (EDC) and N-hydroxy-succinamide (NHS) activating and reducing after, add the coupling of glucose molecule recognition unit phenyl boric acid and make the magnetic microsphere with the visual measuring ability of glucose that glucose molecule is modified, after the phosphate buffer washing, measure the reflectance spectrum of described magnetic microsphere.
(4) glucose molecule detects
After magnetic microsphere usefulness trishydroxymethylaminomethane (Tris) the cushioning liquid washing with visual measuring ability for preparing, in the Tris of 0.5 ml buffer solution, mix 35 ℃ of reaction 1h with tear to be measured.React complete after, with magnet free magnetic microsphere is assembled, according to the color of magnetic microsphere before and after the reaction or by measuring the reflectance spectrum of magnetic microsphere, determine the content of glucose in the tear.
Embodiment 2Use the glucose content in the described magnetic microsphere detection urine
(1) preparation photon crystal micro-ball
The Nano particles of silicon dioxide of monodispersed 200 nm is carried out centrifugal purification, the mass percent of regulating colloidal nanoparticles in the nano-particle solution is 35%, after the ultrasonic dispersion as the water in micro-fluidic, the foreign minister adopts the silicone oil that contains surfactant, collect the drop that produces, after it is heating and curing, puts into Muffle furnace and carry out 800 ℃ of calcinings to improve the stability of photon crystal micro-ball.
(2) preparation magnetic photonic crystal hydrogel microsphere
The photon crystal micro-ball of magnetic photonic crystal hydrogel microsphere preparation in the step (1) is as masterplate, and the method by stencil duplicating obtains.Photon crystal micro-ball is added the Fe that contains light trigger HMPP (0.1%, mass volume ratio) and monodispersed 80nm
3O
4In 15% the polyethyleneglycol diacrylate solution of nano particle, after fully soaking, behind high voltage mercury lamp radiation 1 min, get the magnetic photonic crystal hydrogel microsphere, resulting magnetic photonic crystal hydrogel microsphere is peeled off, washing is placed on 2h in the solvent, obtains the magnetic photonic crystal hydrogel microsphere of counter opal structure.
(3) modification of glucose molecule recognition unit
The magnetic photonic crystal hydrogel microsphere that step (2) is prepared is at the NaOH(10% of 0.1M N, N, N', the N'-tetramethylethylenediamine) in the solution reduction 1h after, wash with 2-(N-morpholine) ethyl sulfonic acid (MES) cushioning liquid, in the MES buffer solution, add 1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimides (EDC) and N-hydroxy-succinamide (NHS) activating and reducing after, add the coupling of glucose molecule recognition unit phenyl boric acid and make the magnetic microsphere with the visual measuring ability of glucose that glucose molecule is modified, after the phosphate buffer washing, measure the reflectance spectrum of magnetic microsphere.
(4) glucose molecule detects
With the magnetic microsphere with the visual measuring ability of glucose for preparing with the phosphate buffer washing after, in the phosphate buffer of 0.5 ml, mixes with urine to be measured, 35 ℃ are reacted 1h.React complete after, with magnet free magnetic microsphere is assembled, according to the color of magnetic microsphere before and after the reaction or by measuring the reflectance spectrum of magnetic microsphere, determine the content of glucose in the urine.
Embodiment 3Use the glucose content in the described magnetic microsphere detection saliva
(1) preparation photon crystal micro-ball
The Nano particles of silicon dioxide of monodispersed 180 nm is carried out centrifugal purification, the mass percent of regulating colloidal nanoparticles in the nano-particle solution is 30%, after the ultrasonic dispersion as the water in micro-fluidic, the foreign minister adopts the hexadecane that contains surfactant, collect the drop that produces, after it is heating and curing, puts into Muffle furnace and carry out 800 ℃ of calcinings to improve the stability of photon crystal micro-ball.
(2) preparation magnetic photonic crystal hydrogel microsphere
The photon crystal micro-ball of magnetic photonic crystal hydrogel microsphere preparation in the step (1) is as masterplate, and the method by stencil duplicating obtains.The photon crystal micro-ball adding is contained light trigger HMPP (0.1%, mass volume ratio) and in the polyethyleneglycol diacrylate and acrylic acid solution of the Ni nano particle of monodispersed 100nm, after fully soaking, behind high voltage mercury lamp radiation 8 min, get the magnetic photonic crystal hydrogel microsphere, resulting magnetic photonic crystal hydrogel microsphere is peeled off, washing is placed on 2 h in the solvent, obtains the magnetic photonic crystal hydrogel microsphere of counter opal structure.
(3) modification of glucose molecule recognition unit
The magnetic photonic crystal hydrogel microsphere that step (2) is prepared is at the NaOH(10% of 0.1M N, N, N', the N'-tetramethylethylenediamine) in the solution reduction 1h after, wash with 2-(N-morpholine) ethyl sulfonic acid (MES) cushioning liquid, in the MES buffer solution, add 1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimides (EDC) and N-hydroxy-succinamide (NHS) activating and reducing after, add the coupling of glucose molecule recognition unit phenyl boric acid and make the magnetic microsphere with the visual measuring ability of glucose that glucose molecule is modified, after the phosphate buffer washing, measure the reflectance spectrum of magnetic microsphere.
(4) glucose molecule detects
With the magnetic microsphere with the visual measuring ability of glucose for preparing with the phosphate buffer washing after, in the phosphate buffer of 0.5 ml, mixes with saliva to be measured, 35 ℃ are reacted 0.5 h.React complete after, with magnet free magnetic microsphere is assembled, according to the color of magnetic microsphere before and after the reaction or by measuring the reflectance spectrum of magnetic microsphere, determine the content of glucose in the saliva.
Embodiment 4Use the glucose content in the described magnetic microsphere detection sweat
(1) preparation photon crystal micro-ball
The polystyrene nanoparticle of monodispersed 160 nm is carried out centrifugal purification, the mass percent of regulating colloidal nanoparticles in the nano-particle solution is 50%, after the ultrasonic dispersion as the water in micro-fluidic, the foreign minister adopts the hexadecane that contains surfactant, collect the drop that produces, after it is heating and curing, puts into Muffle furnace and carry out 800 ℃ of calcinings to improve the stability of photon crystal micro-ball.
(2) preparation magnetic photonic crystal hydrogel microsphere
The photon crystal micro-ball of magnetic photonic crystal hydrogel microsphere preparation in the step (1) is as masterplate, and the method by stencil duplicating obtains.The photon crystal micro-ball adding is contained thermal initiator ammonium persulfate, N, N, N', N'-tetramethylethylenediamine (0.1%, mass volume ratio) (29:1 and in the mixed solution of the NIPA of the Ni nano particle of monodispersed 80nm and methylene diacrylamide, mol ratio), after fully soaking, behind polymerized at room temperature 30 min, get the magnetic photonic crystal hydrogel microsphere, resulting magnetic photonic crystal hydrogel microsphere is peeled off, washing is placed on 2 h in the solvent, obtains the magnetic photonic crystal hydrogel microsphere of counter opal structure.
(3) modification of glucose molecule recognition unit
The photonic crystal hydrogel microsphere that second step is prepared is at the NaOH(10% of 0.1 M N, N, N', the N'-tetramethylethylenediamine) in the solution reduction 1h after, wash with 2-(N-morpholine) ethyl sulfonic acid (MES) cushioning liquid, in the MES buffer solution, add 1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimides (EDC) and N-hydroxy-succinamide (NHS) activating and reducing after, add the coupling of glucose molecule recognition unit phenyl boric acid and make the magnetic microsphere with the visual measuring ability of glucose that glucose molecule is modified, after the phosphate buffer washing, measure the reflectance spectrum of magnetic microsphere.
(4) glucose molecule detects
After magnetic microsphere usefulness trishydroxymethylaminomethane (Tris) the cushioning liquid washing for preparing, in the Tris of 0.5 ml buffer solution, mix 35 ℃ of reaction 1 h with sweat to be measured.React complete after, with magnet free magnetic microsphere is assembled, according to the color of magnetic microsphere before and after the reaction or by measuring the reflectance spectrum of magnetic microsphere, determine the content of glucose in the sweat.
Claims (8)
1. the preparation method with magnetic microsphere of the visual measuring ability of glucose is characterized in that, described magnetic microsphere prepares by following steps:
(1) preparation process of magnetic photonic crystal hydrogel microsphere:
Photon crystal micro-ball is added before the hydrogel contain magnetic nano-particle in the aggressiveness solution, add again initator, aggressiveness solution is cured before curing reaction is to hydrogel, obtain the magnetic photonic crystal hydrogel microsphere, the described magnetic photonic crystal hydrogel microsphere that obtains is peeled off, place solvent to remove the photon crystal micro-ball masterplate after the washing, take out after two hours with the pure water washing, then be kept in the pure water;
(2) have the preparation process of the magnetic microsphere of the visual measuring ability of glucose:
Method by the chemical bond coupling, glucose molecule recognition unit phenyl boric acid is coupled on the magnetic photonic crystal hydrogel microsphere that makes in the step (1), make the magnetic microsphere with the visual measuring ability of glucose, the described magnetic microsphere with the visual measuring ability of glucose for preparing with after the buffer solution washing, is measured its reflectance spectrum.
2. the preparation method with magnetic microsphere of the visual measuring ability of glucose according to claim 1, it is characterized in that, described photon crystal micro-ball is assembled by Nano particles of silicon dioxide, poly-methyl acid methyl esters nano particle or polystyrene nanoparticle.
3. the preparation method with magnetic microsphere of the visual measuring ability of glucose according to claim 1, it is characterized in that, aggressiveness is any one or multiple the mixing in acrylamide, methyl methacrylate, poly hydroxy ethyl acrylate, acetylbutyrylcellulose, siloxanes methacrylate, fluorine silicon methacrylate, perfluor ether, N-vinylpyrrolidone, polyvinyl alcohol, GMA or the GDMA before the described hydrogel.
4. the preparation method with magnetic microsphere of the visual measuring ability of glucose according to claim 1 is characterized in that, described magnetic photonic crystal hydrogel microsphere has counter opal structure.
5. the preparation method with magnetic microsphere of the visual measuring ability of glucose according to claim 1 is characterized in that, the method for described curing reaction is specially the UV-irradiation polymerization or uses baking oven to carry out heat cure.
6. the preparation method with magnetic microsphere of the visual measuring ability of glucose according to claim 1 is characterized in that, described chemical bond coupling method comprises amino with amino reaction, amino and the reaction of carboxyl or the reaction of carboxyl and hydroxyl.
7. the method for the visual detection of glucose is characterized in that:
According to claim 1, the magnetic microsphere with the visual measuring ability of glucose made to one of 6 described preparation methods mixed with glucose solution in cushioning liquid carry out the specific binding reaction, react complete after, utilize magnet with free magnetic microsphere polymerization, relatively react the color of front and back magnetic microsphere or by measuring the reflectance spectrum of magnetic microsphere, determine the content of glucose.
8. the method for the visual detection of glucose according to claim 7 is characterized in that, described glucose solution is tear, urine, saliva or sweat.
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| CN111440355B (en) * | 2020-04-17 | 2020-12-08 | 南京鼓楼医院 | Preparation method and application of magnetic structure color hydrogel microcarrier for bladder cancer protein multivariate analysis |
| CN111504941A (en) * | 2020-05-14 | 2020-08-07 | 中国人民解放军陆军军医大学第一附属医院 | Terahertz attenuated total reflection technology platform for unmarked evaluation of response characteristics of responsive hydrogel |
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