Methacrylic acid inverse opal colloidal crystal microsphere as well as preparation method and application thereof
By forming a methacrylic gel network in the gaps of silica photonic crystal microspheres, inverse opal colloidal crystal microspheres were prepared, which solved the complex and expensive problems of the existing cisplatin detection methods, and achieved simple, fast and high-accuracy detection effects.
Patent Information
- Application Number
- CN202411614750.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-11-13
- Publication Date
- 2025-03-04
AI Technical Summary
The existing cisplatin detection methods have complex sample preprocessing, expensive equipment costs and cumbersome operation problems, making it difficult to achieve simple, cheap, fast and accurate detection.
By chemical crosslinking in the gaps of silica photonic crystal microspheres, a stable methacrylic gel network was formed, and the silica structure was subsequently corroded to obtain inverse opal colloidal crystal microspheres based on methacrylic gels, which were used to detect cisplatin molecules.
It realizes simple, fast and highly selective detection of cisplatin, which is easy to operate, intuitive and easy to understand, and can improve detection accuracy in complex environments.
Smart Images
Figure CN119553370A_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the field of detection technology, relates to a detection application of cisplatin molecules, and in particular to a methacrylic acid inverse opal colloidal crystal microsphere and a preparation method and application thereof. Background Art
[0002] The anti-tumor drug cisplatin is a widely used platinum-based chemotherapy drug. With its efficient anti-tumor activity, it plays an important role in the treatment of various solid tumors (such as testicular cancer, ovarian cancer, lung cancer, bladder cancer, and head and neck cancer). Cisplatin inhibits the growth and division of tumor cells by causing DNA inter-chain and intra-chain crosslinks, interfering with DNA replication and transcription. However, the clinical application of cisplatin is also accompanied by significant toxicity and side effects, including severe nephrotoxicity, ototoxicity, neurotoxicity, and gastrointestinal reactions. These adverse reactions not only limit the dosage and course of cisplatin, but may also affect the patient's quality of life and even threaten his life. In order to ensure the safety and effectiveness of cisplatin treatment, it is particularly important to accurately monitor its concentration in the body. At present, a variety of methods have been developed for the detection of cisplatin, including high performance liquid chromatography (HPLC), inductively coupled plasma mass spectrometry (ICP-MS), electrochemical sensors, and fluorescent probes. Although the existing detection methods have made significant progress in sensitivity and accuracy, problems such as complex sample pretreatment, expensive equipment costs, and cumbersome operations still exist. Therefore, developing a simple, cheap, rapid and accurate method for detecting cisplatin has become a research hotspot. Summary of the invention
[0003] In view of the deficiencies in the prior art, the present invention provides a methacrylic acid inverse opal colloidal crystal microsphere, a preparation method and an application thereof. A pre-gel solution containing methacrylic acid is chemically cross-linked in the gaps within a silica photonic crystal microsphere to form a stable methacrylic acid gel network, and then the inverse opal colloidal crystal microsphere based on methacrylic acid gel is obtained by corroding the silica structure. The synthesis steps are simple, and the microsphere is used to detect cisplatin molecules with the characteristics of simple, rapid and high selectivity.
[0004] To achieve the above object, the present invention adopts the following technical solutions:
[0005] A method for preparing methacrylic acid inverse opal colloidal crystal microspheres comprises the following steps: step 1, preparing silicon dioxide photonic crystal microspheres; step 2, placing the silicon dioxide photonic crystal microspheres in a pre-gel solution containing methacrylic acid and fully immersing them under light-proof conditions to ensure that the pre-gel solution containing methacrylic acid fully penetrates into the gaps inside the silicon dioxide photonic crystal microspheres; then inducing the pre-gel solution containing methacrylic acid to undergo chemical cross-linking and curing in the gaps inside the silicon dioxide photonic crystal microspheres to form a stable methacrylic acid gel network; and corroding silicon dioxide to obtain inverse opal colloidal crystal microspheres based on methacrylic acid gel.
[0006] To optimize the above technical solutions, the specific measures taken also include:
[0007] Furthermore, in step 1, the preparation method of the silica photonic crystal microspheres is: emulsifying, drying, washing and calcining silica nanoparticles in sequence to obtain silica photonic crystal microspheres. The particle size of the silica photonic crystal microspheres is 200-300 μm.
[0008] Furthermore, in step one, the emulsification method is: the silica nanoparticles are configured into a silica nanoparticle aqueous solution, the silica nanoparticle aqueous solution is used as the internal phase solution, and dimethyl silicone oil is used as the external phase solution, and silica nanoparticle droplets are prepared by a microfluidic device; the flow rates of the internal phase solution and the external phase solution are 1-3 ml / h and 5-8 ml / h, respectively.
[0009] Furthermore, in step 1, the drying method is: first drying at 60-70°C for 12-18 hours, and then drying at 80-90°C for 2-3 hours.
[0010] Furthermore, in step 1, the calcination method is: firstly raising the temperature from room temperature to 800-900°C over 3-4 hours and continuing the calcination for 4-5 hours, and then cooling from 800-900°C to room temperature over 3-4 hours.
[0011] Furthermore, in step 2, ultraviolet light is used to induce the curing of the pre-gel solution containing methacrylic acid; the pre-gel solution containing methacrylic acid includes methacrylic acid (mAA), polyethylene glycol diacrylate (PEGDA), acrylamide / N, N-dimethylbispropylamide (AAm / Bis), water and 2-hydroxy-2-methyl-1-phenyl-1-propanone; wherein PEGDA can improve the mechanical strength of the hydrogel; AAm / Bis can increase the dissolution of mAA and avoid the problem of uneven dissolution of mAA; and The volume ratio of acrylic acid, polyethylene glycol diacrylate, acrylamide / N,N-dimethylbispropylamide, water and 2-hydroxy-2-methyl-1-phenyl-1-propanone is 2-7:0-10:25-75:10-60:1-2, preferably 2-7:8:45:38-43:2; in acrylamide / N,N-dimethylbispropylamide, the mass ratio of acrylamide and N,N-dimethylbispropylamide is 29:1; the conditions of ultraviolet irradiation are: wavelength of 365nm, crosslinking time of 10-60s.
[0012] Furthermore, in step 2, the soaking time is 0.2 to 3 hours.
[0013] Furthermore, in step 2, silicon dioxide is etched using a hydrofluoric acid solution, the mass volume ratio concentration of the hydrofluoric acid solution is 1 to 8%, and the etching time is 0.5 to 3 hours.
[0014] The invention also provides methacrylic acid inverse opal colloidal crystal microspheres prepared by the preparation method.
[0015] The present invention also provides application of the methacrylic acid inverse opal colloidal crystal microspheres in detecting cisplatin molecules.
[0016] The beneficial effects of the present invention are:
[0017] The present invention provides a methacrylic acid inverse opal colloidal crystal microsphere, a preparation method and an application thereof, which combines the photonic bandgap effect of photonic crystals and the specific interaction between methacrylic acid and cisplatin, thereby realizing a simple and quick detection of cisplatin. Specifically, cisplatin, as a metal complex, has two amino ligands and two chloride ion ligands, and its combination with methacrylic acid (mAA) mainly depends on the synergistic effect of various chemical forces such as coordination bonds, electrostatic interactions and hydrogen bonds. The coordination bonds are formed by the Pt^2+ metal center of cisplatin and the oxygen atom in the carboxyl group of mAA, the electrostatic interaction occurs between the positive charge part generated by the hydrolysis of cisplatin and the mAA anion, and the hydrogen bond further stabilizes the combination through the interaction between the mAA carboxyl group and the amino ligand of cisplatin. In addition, the inverse opal structure is a material with a three-dimensional ordered periodic structure, which can produce structural colors, and provides the possibility for the visual detection of cisplatin. By observing the changes in the color of the inverse opal structure, the presence or absence of cisplatin can be preliminarily judged, and this method is simple to operate, intuitive and easy to understand. At the same time, the structure is selective in the diffraction of light. By adjusting the structural parameters, the detection effect of cisplatin can be optimized and the detection accuracy in complex environments can be improved. BRIEF DESCRIPTION OF THE DRAWINGS
[0018] Figure 1 It is a schematic diagram of the synthesis route of methacrylate inverse opal colloidal crystal microspheres;
[0019] Figure 2 is a scanning electron microscope image of the silicon dioxide photonic crystal microsphere of Example 1;
[0020] Figure 3 is a scanning electron microscope image of the methacrylic acid inverse opal colloidal crystal microspheres of Example 1;
[0021] Figure 4 This is a graph showing the spectrum changes of the methacrylic acid inverse opal colloidal crystal microspheres in Example 1 in cisplatin solutions of different concentrations;
[0022] Figure 5 It is a statistical graph of the spectrum shift of the methacrylic acid inverse opal colloidal crystal microspheres of Example 1 in different interfering ion solutions. DETAILED DESCRIPTION
[0023] The present invention is further described below in conjunction with specific embodiments.
[0024] Example 1
[0025] This embodiment provides a methacrylic acid inverse opal colloidal crystal microsphere for detecting cisplatin molecules, and the preparation method thereof is as follows: Figure 1 As shown, the following steps are included:
[0026] Step 1: Synthesize silica photonic crystal microspheres:
[0027] A 20% (w / v) aqueous solution of silica nanoparticles was loaded into a syringe as the inner phase solution, and a syringe filled with dimethyl silicone oil (50 cs) was used as the outer phase solution. The two syringes were then fixed on a microfluidic pump (LSP01-1A, Baoding Lange Constant Flow Pump Co., Ltd.). The inner phase solution and the outer phase solution were respectively introduced into the inner phase port and the outer phase port of a PE capillary microfluidic device through the microfluidic pump. The flow rate was controlled by the microfluidic device (1.25 mL / h for the inner phase solution and 7.5 mL / h for the outer phase solution), and microdroplets of uniform size, i.e., silica nanoparticle emulsion droplets, were obtained at the confluence of the two phase outlets.
[0028] The collected silica nanoparticle droplets were placed in a 65°C oven overnight (16 hours), and the temperature was adjusted to 85°C and maintained for 2 hours the next day to ensure complete evaporation of water. The excess dimethyl silicone oil on the droplets was then removed by repeated washing with n-hexane, and the droplets were collected in a crucible and washed five times to ensure that there was no residual silicone oil. Finally, the washed droplets were transferred to a muffle furnace for calcination, first from 30°C to 800°C after 3 hours and continued calcination for 4 hours, and then from 800°C to 30°C after 3 hours. After calcination, solid silica photonic crystal microspheres were obtained.
[0029] The results of scanning electron microscopy analysis of silica photonic crystal microspheres are as follows Figure 2 As shown, the microspheres are self-assembled from multiple nanoparticles of uniform size, and the stacking morphology is a microsphere with a particle size of 280 μm. The surface and the interior have three-dimensional interconnected nano-network gaps, and the porous structure can provide a larger specific surface area.
[0030] Step 2: Preparation of methacrylic acid inverse opal colloidal crystal microspheres:
[0031] A pre-gel solution containing methacrylic acid was prepared, including methacrylic acid (mAA), polyethylene glycol diacrylate (PEGDA), acrylamide / N, N-dimethylbispropylamide (AAm / Bis, the mass ratio of acrylamide and N, N-dimethylbispropylamide was 29:1), water and 2-hydroxy-2-methyl-1-phenyl-1-propanone, and the volume ratio was 3:8:45:42:2. Then the prepared silica photonic crystal microspheres were divided into multiple groups, each with 10 microspheres, and each group of microspheres was added to a 200 μL tube, and 100 μL of the pre-gel solution containing methacrylic acid was added to the tube under light-proof conditions and soaked for 2 hours. Subsequently, ultraviolet light with a wavelength of 365nm was irradiated for 20s to induce chemical cross-linking and curing of the pre-gel solution containing methacrylic acid in the gaps inside the microspheres. The obtained hydrogel microspheres were immersed in water, and the residual hydrogel on the surface was peeled off. The microspheres were then corroded by 4% (w / v) hydrofluoric acid solution for 2h to etch away the silica structure, thereby obtaining inverse opal colloidal crystal microspheres based on methacrylic acid gel.
[0032] The results of scanning electron microscopy analysis of methacrylate inverse opal colloidal crystal microspheres are as follows: Figure 3 As shown, the pores of the microspheres are filled with methacrylic acid pre-gel solution.
[0033] Example 2
[0034] This embodiment provides a methacrylic acid inverse opal colloidal crystal microsphere for detecting cisplatin molecules. The preparation method is basically the same as that in the embodiment, the only difference is that in the pre-gel solution containing methacrylic acid, methacrylic acid (mAA), polyethylene glycol diacrylate (PEGDA), acrylamide / N,N-dimethylbispropylamide (AAm / Bis), water and 2-hydroxy-2-methyl-1-phenyl-1-propanone are in a volume ratio of 4:8:45:41:2.
[0035] Example 3
[0036] This embodiment provides a methacrylic acid inverse opal colloidal crystal microsphere for detecting cisplatin molecules. The preparation method is basically the same as that in the embodiment, the only difference is that in the pre-gel solution containing methacrylic acid, methacrylic acid (mAA), polyethylene glycol diacrylate (PEGDA), acrylamide / N,N-dimethylbispropylamide (AAm / Bis), water and 2-hydroxy-2-methyl-1-phenyl-1-propanone are in a volume ratio of 5:8:45:40:2.
[0037] The relevant performance characteristics of the methacrylic acid inverse opal colloidal crystal microspheres used in Examples 1 to 3 for detecting cisplatin molecules are as follows:
[0038] Characterization of the sensitivity of methacrylate inverse opal colloidal crystal microspheres to cisplatin molecules:
[0039] 100 μL of cisplatin test solutions of different concentrations (0, 0.001, 0.01, 0.1, 1, 10, 100, 1000 μg / ml) were prepared (the solvent was normal saline), and the reflection peak before the reaction was recorded as F0. The cisplatin test solutions of different concentrations were added to 8 200 μL tubes, respectively, and then 10 methacrylate inverse opal colloidal crystal microspheres of Example 1 were added respectively, and the reflection peak after 2 hours of reaction was recorded as F, and the reflection peak offset before and after the reaction was calculated.
[0040] The spectral changes of methacrylate inverse opal colloidal crystal microspheres to different concentrations of cisplatin solutions are shown in Figure 2. Figure 4 As shown in the figure, with the gradual increase of cisplatin concentration, the diffraction peak of the inverse opal colloidal crystal microspheres gradually red-shifted and showed regular changes. When the concentration reached above 100 μg / ml, the reflection peak offset reached more than 23 nm.
[0041] The above method was used to test the spectral shift of the methacrylic acid inverse opal colloidal crystal microspheres of Examples 2 and 3. The methacrylic acid concentration of Example 3 was high, and the spectral shift was more obvious.
[0042] Characterization of the Selective Recognition of Cisplatin Molecules by Methacrylate Inverse Opal Colloidal Crystal Microspheres:
[0043] Prepare 100 μg / ml of different ions (K + , Li + , Cs + , Ca 2+ , Pb 2+ Mg 2+ 、Al 3+ , Cu 2+ , OHP) test solution, record the reflection peak before the reaction as λ0, add different ion test solutions with a concentration of 100 μg / ml into 9 200 μL tubes, and then add 10 methacrylic acid photonic crystal microspheres of Example 1, respectively, record the reflection peak after 2 hours of reaction as λ, and calculate the reflection peak offset before and after the reaction.
[0044] The reflection peak shift of methacrylate inverse opal colloidal crystal microspheres to different ion solutions is shown in the figure. Figure 5 As shown in Figure 2, these ions have little effect on the reflection peak shift of methacrylate inverse opal colloidal crystal microspheres, all less than 5 nm, while the same concentration of cisplatin solution can cause a shift of about 23 nm (see Figure 4 ), this comparison shows that methacrylic acid inverse opal colloidal crystal microspheres have high specific recognition ability for cisplatin molecules.
[0045] In the present invention, unless otherwise specified, the scientific and technical terms used herein have the meanings commonly understood by those skilled in the art. In addition, the reagents, materials and operating procedures used herein are reagents, materials and conventional procedures widely used in the corresponding fields.
[0046] Finally, it should be noted that the above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art can still modify the technical solutions described in the aforementioned embodiments or replace some of the technical features therein by equivalents. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.
Claims
1. A method for preparing methacrylic acid inverse opal colloidal crystal microspheres, characterized in that: The following steps are involved: Step 1, preparing silicon dioxide photonic crystal microspheres; Step 2: Under light-proof conditions, fully immerse the silica photonic crystal microspheres in a pre-gel solution containing methacrylic acid; then induce the pre-gel solution containing methacrylic acid to undergo chemical cross-linking and solidification in the internal gaps of the silica photonic crystal microspheres to form a stable methacrylic acid gel network; and corrode the silica to obtain inverse opal colloidal crystal microspheres based on methacrylic acid gel.
2. The method for preparing methacrylic acid inverse opal colloidal crystal microspheres according to claim 1, characterized in that: In step 1, the method for preparing the silica photonic crystal microspheres is: emulsifying, drying, washing and calcining silica nanoparticles in sequence to obtain silica photonic crystal microspheres.
3. The method for preparing methacrylic acid inverse opal colloidal crystal microspheres according to claim 2, characterized in that: In step 1, the emulsification method is: the silica nanoparticles are configured into a silica nanoparticle aqueous solution, the silica nanoparticle aqueous solution is used as the internal phase solution, and dimethyl silicone oil is used as the external phase solution, and the silica nanoparticle emulsion droplets are prepared by a microfluidic device; The flow rates of the inner phase solution and the outer phase solution are 1-3 ml / h and 5-8 ml / h respectively.
4. The method for preparing methacrylic acid inverse opal colloidal crystal microspheres according to claim 2, characterized in that: In step 1, the drying method is: first drying at 60-70° C. for 12-18 hours, and then drying at 80-90° C. for 2-3 hours.
5. The method for preparing methacrylic acid inverse opal colloidal crystal microspheres according to claim 2, characterized in that: In step 1, the calcination method is: firstly, heating from room temperature to 800-900°C over 3-4 hours and continuously calcining for 4-5 hours, and then cooling from 800-900°C to room temperature over 3-4 hours.
6. The method for preparing methacrylic acid inverse opal colloidal crystal microspheres according to claim 1, characterized in that: In step 2, ultraviolet light is used to induce the pre-gel solution containing methacrylic acid to solidify; The pre-gel solution containing methacrylic acid comprises methacrylic acid, polyethylene glycol diacrylate, acrylamide / N,N-dimethylbispropylamide, water and 2-hydroxy-2-methyl-1-phenyl-1-propanone, and the volume ratio thereof is 2-7:0-10:25-75:10-60:1-2; in acrylamide / N,N-dimethylbispropylamide, the mass ratio of acrylamide to N,N-dimethylbispropylamide is 29:1; The conditions of ultraviolet irradiation are: wavelength of 365 nm, and cross-linking time of 10 to 60 s.
7. The method for preparing methacrylic acid inverse opal colloidal crystal microspheres according to claim 6, characterized in that: In step 2, the soaking time is 0.2 to 3 hours.
8. The method for preparing methacrylic acid inverse opal colloidal crystal microspheres according to claim 1, characterized in that: In step 2, silicon dioxide is etched using a hydrofluoric acid solution, the mass volume ratio concentration of the hydrofluoric acid solution is 1 to 8%, and the etching time is 0.5 to 3 hours.
9. Methacrylic acid inverse opal colloidal crystal microspheres prepared by the preparation method according to any one of claims 1 to 8.
10. Use of the methacrylic acid inverse opal colloidal crystal microspheres as claimed in claim 1 in detecting cisplatin molecules.