CN110018142B - Composite fluorescent substrate, preparation method and application of composite fluorescent substrate - Google Patents
Composite fluorescent substrate, preparation method and application of composite fluorescent substrate Download PDFInfo
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Abstract
本发明公开了复合荧光基底、复合荧光基底的制备方法以及应用。该复合荧光基底包括凝胶和附着在所述凝胶的孔隙表面的分子印迹聚合物材料。凝胶具有三维网状结构，能够提供较大的比表面积来附着MIP材料；由于基底具有凝胶，凝胶具有粘性且为固状，当凝胶与MIP材料协同作用时，不仅能够通过切取的方式随取随用，而且还能通过擦拭待测样品表面的方式快速富集目标物，并利用固体表面荧光可直接测定固体样品的优势，快速取样和测试，与现有技术相比，无需对样品进行复杂的预处理即可实现对目标物的快速检测。
The invention discloses a composite fluorescent substrate, a preparation method and application of the composite fluorescent substrate. The composite fluorescent substrate includes a gel and a molecularly imprinted polymer material attached to the pore surface of the gel. The gel has a three-dimensional network structure, which can provide a large specific surface area to attach the MIP material; because the substrate has a gel, the gel is viscous and solid, and when the gel cooperates with the MIP material, it can not only pass the excised The method can be used at any time, and the target can be quickly enriched by wiping the surface of the sample to be tested, and the advantage of using the solid surface fluorescence to directly measure the solid sample, fast sampling and testing, compared with the existing technology, no need for The complex pretreatment of the sample can realize the rapid detection of the target.
Description
Technical Field
The invention relates to the technical field of detection of illegal additives in food, in particular to the technical field of detection of rhodamine B, and specifically relates to a composite fluorescent substrate, a preparation method of the composite fluorescent substrate and application of the composite fluorescent substrate.
Background
The instrumental analysis techniques used for the detection of illegal additives in food nowadays mainly include spectroscopic techniques including ultraviolet-visible absorption spectroscopy (μ V-Vis), Atomic Absorption Spectroscopy (AAS), etc.; the most common chromatographic techniques used are high performance liquid chromatography, gas chromatography-mass spectrometry (GC-MS); the main application of electrochemical technology is high efficiency Capillary Electrophoresis (CE). The precision of the methods can meet the requirements of experiments, the lowest detection limit can also meet the requirements of trace analysis, but the methods have the problems of narrow application range, expensive equipment, complex operation, high cost and the like; moreover, most of chromatographic analyses need special detection instruments, and have high requirements on detection personnel, so that the chromatographic analyses are not suitable for large-scale field rapid detection. In the aspect of rapid detection, a rapid detection method developed according to the principle of solid-phase extraction is adopted, but the method mainly adopts the visual observation of a red strip on a solid-phase extraction column for judgment, so that false positive is easy to occur, the accuracy is low, and the detection sensitivity is not high. Therefore, it is important to establish a novel method for rapidly and reliably detecting illegal additives in food.
A Molecular Imprinted Polymer (MIP) is a Polymer with specific recognition and selective adsorption. Molecular Imprinting Solid Phase Extraction (MISPE) is a novel sample pretreatment technology developed in recent years, and a Solid Phase Extraction material is prepared by utilizing a specific selective adsorption mechanism of MIP.
When the MIP is used for detecting the target object, firstly, the sample needs to be pretreated, namely, the target object on the surface of the sample is dissolved by a solvent, the organic liquid containing the target object is obtained by filtering and centrifuging and then extracting, and then the organic liquid passes through a solid phase flow column filled with the MIP material, so that the target object in the organic liquid is adsorbed by the MIP material and then the MIP material is detected. Therefore, the detection of the target object by using the MIP requires a large amount of pretreatment time, and the pretreatment process is complex in operation and poor in reproducibility.
Disclosure of Invention
The invention mainly aims to provide a composite fluorescent substrate, a preparation method and application of the composite fluorescent substrate, and aims to solve the technical problems that in the prior art, when a molecularly imprinted polymer is used for testing a target object in a sample, the sample is subjected to overlong pretreatment time and the pretreatment process is complicated.
In order to achieve the above object, according to one aspect of the present invention, there is provided a composite fluorescent substrate. The composite fluorescent substrate comprises gel and molecularly imprinted polymer material attached to the pore surfaces of the gel.
Firstly, the composite fluorescent substrate is used for solid surface fluorescence spectrum detection, and has the advantages of simplicity, rapidness and high sensitivity. Secondly, the defect that which substance is wide in fluorescence spectrum range and cannot be accurately determined is overcome by utilizing the characteristic of strong specificity of MIP materials, and more enrichment can be carried out on the sample to be detected in the actual detection process. Moreover, the anti-interference performance is stronger, can carry out the qualitative detection of trace to the sample that awaits measuring fast. Meanwhile, the gel has a three-dimensional network structure and can provide a larger specific surface area for attaching the MIP material; because the substrate has the gel, and the gel has viscidity and is solid, when gel and MIP material synergistic effect, not only can be through the mode of cutting and take along with using, but also can be through the mode of wiping the sample surface that awaits measuring enrichment target thing fast to utilize the advantage that solid surface fluorescence can directly determine the solid sample, quick sample and test, compare with prior art, need not to carry out complicated preliminary treatment to the sample and can realize the quick detection to the target thing.
Further, the gel is a PVA gel; the molecularly imprinted polymer is rhodamine B molecularly imprinted polymer. The PVA (polyvinyl alcohol) gel is easy to obtain and has a stable state, so that the rapid detection of rhodamine B (shown as RhB, the same below) is realized.
Further, the mass ratio of the rhodamine B molecularly imprinted polymer to the PVA is (0.05-1): (0.2-1); sodium tetraborate (molecular formula: Na)2B4O7·10H2O). The stability of PVA gel may be influenced by the excessive content of rhodamine B molecularly imprinted polymerSex; if the rhodamine B molecularly imprinted polymer is too low, the target substance enrichment capacity is poor. Therefore, the rhodamine B molecularly imprinted polymer is uniformly distributed in the PVA gel, and the obtained composite fluorescent substrate has strong enrichment capacity on the target. The sodium tetraborate is used as the coagulant aid, and the stable PVA gel can be quickly obtained.
Furthermore, the mass ratio of PVA to sodium tetraborate is (1-5): 0.4-2. According to verification, when the mass ratio of PVA to sodium tetraborate is (1-5) to (0.4-2), the fluorescence signal of the obtained composite fluorescent substrate is the best.
In order to achieve the above object, according to another aspect of the present invention, there is also provided a method for preparing a composite fluorescent substrate, comprising the steps of:
(1) obtaining a mixed solution containing a gel precursor and a molecularly imprinted polymer material;
(2) adding a coagulant aid for gelling the gel precursor into gel, and stirring until the gel is formed, thereby obtaining the composite fluorescent substrate.
The preparation method of the composite fluorescent substrate has the advantages of simple process, short synthesis time, good stability of the obtained composite fluorescent substrate, proper hardness and viscosity, rapid sampling and testing, and rapid detection of the target without complex pretreatment of the sample.
Further, the gel is PVA gel, and the coagulant aid is sodium tetraborate; the molecularly imprinted polymer is rhodamine B molecularly imprinted polymer.
Further, the step (1) specifically comprises: firstly, obtaining a PVA solution, and then adding a rhodamine B molecularly imprinted polymer into the PVA solution; the mass fraction of PVA in the PVA solution is 4-10%; the PVA in the mixed solution is fully dissolved and uniformly dispersed to help the PVA gel, and compared with the process of directly blending the PVA and the molecularly imprinted polymer material, the PVA solution is firstly obtained, and then the molecularly imprinted polymer is dispersed, so that the influence of the molecularly imprinted polymer on the PVA solubility can be avoided, and the stable gel can be quickly obtained.
Further, the sodium tetraborate is added into the PVA solution after being prepared into the solution, and the mass ratio of the PVA to the sodium tetraborate is (1-5): (0.4-2). By adding dissolved sodium tetraborate, the sodium tetraborate can be quickly and uniformly distributed in the PVA solution, which is beneficial to quickly obtaining a substrate with consistent performance at each position.
Further, the rhodamine B molecularly imprinted polymer is prepared from SiO2Is carrier, methacrylic acid (MAA, the same below) is functional monomer, ethylene glycol dimethacrylate (EGDMA, the same below) is cross-linking agent, acetonitrile is pore-forming agent and 2, 2' -azobisisobutyronitrile (AIBN, the same below) is initiator. Therefore, the Surface molecularly Imprinted Polymer material (Surface Molecular Imprinted Polymer, SMIP for short) with the Imprinted sites obviously higher than that of the traditional molecularly Imprinted Polymer is obtained.
In order to achieve the above object, according to another aspect of the present invention, there is also provided a method for detecting rhodamine B, comprising the steps of:
(1) cutting a composite fluorescent substrate, wherein the composite fluorescent substrate is the composite fluorescent substrate or the composite fluorescent substrate prepared by the preparation method, and the molecularly imprinted polymer is rhodamine B molecularly imprinted polymer;
(2) wiping the surface of the sample to be detected with the cut composite fluorescent substrate;
(3) and carrying out solid surface fluorescence detection on the wiped composite fluorescent substrate to obtain the concentration of rhodamine B on the surface of the sample to be detected.
Therefore, the composite fluorescent substrate or the fluorescent substrate prepared by the preparation method can quickly enrich the target object in the sample to be detected in a wiping mode, and then the concentration of the target object on the surface of the sample to be detected can be quickly detected through the hometown fluorescent spectrum.
Further, the specific process of the wiping is as follows: firstly, spraying a solvent on the surface of a sample to be detected, and then wiping the composite fluorescent substrate on the surface of the sample to be detected through dust-free paper. Therefore, the target substance can be quickly enriched, and the influence of impurities on the test result can be avoided.
Therefore, the composite fluorescent substrate, the preparation method and the application of the composite fluorescent substrate can realize rapid sampling and testing, and the rapid detection of the target object can be realized without performing complex pretreatment on the sample.
The invention is further described with reference to the following figures and detailed description. Additional aspects and advantages of the invention will be set forth in part in the description which follows and, in part, will be obvious from the description, or may be learned by practice of the invention.
Drawings
The accompanying drawings, which are incorporated in and constitute a part of this specification, are included to assist in understanding the invention, and are included to explain the invention and their equivalents and not limit it unduly. In the drawings:
FIG. 1 shows the fluorescence test results of the composite fluorescent substrates of examples 1-5 (designated as RhB-SMIPs-PVA gel substrates, the same applies below).
FIG. 2 shows the results of fluorescence measurements on RhB-SMIPs-PVA gel substrates of examples 3 and 6 to 8.
FIG. 3 shows the results of fluorescence measurements of RhB-SMIPs-PVA gel substrates of examples 3 and 9 to 12.
FIG. 4 shows the fluorescence test results of the RhB-SMIPs-PVA gel substrate of example 3 for different concentrations of RhB solution, wherein curves a-k correspond to RhB solutions of 10mg/L, 8mg/L, 6mg/L, 4mg/L, 2mg/L, 1mg/L, 0.8mg/L, 0.6mg/L, 0.4mg/L, 0.2mg/L and 0.1mg/L, respectively.
FIG. 5 shows the results of fluorescence measurements of RhB-SMIPs-PVA gel substrates of example 3 and PVA gel substrates of comparative example 1 on RhB solutions of different concentrations.
FIG. 6 is the results of fluorescence measurements of RhB-SMIPs-PVA gel substrates of example 3 and PVA gel substrates of control 1 on solutions of Rh6G at different concentrations.
FIG. 7 shows the results of fluorescence measurements of RhB-SMIPs-PVA gel substrates of example 3 for different pigments.
FIG. 8 is the results of detection of RhB on the surface of a pepper powder by the RhB-SMIPs-PVA gel base of example 3, wherein curves a-e are respectively 0.5mg/LRhB soaked pepper powder, 1mg/LRhB soaked pepper powder, 5mg/LRhB soaked pepper powder, 10mg/LRhB soaked pepper powder, and un-soaked pepper powder.
FIG. 9 is SiO2SEM photograph of KH570 with a partially enlarged inset.
FIG. 10 is an SEM photograph of RhB-SMIPs before elution.
FIG. 11 is an SEM photograph of RhB-SMIPs after washing away the template molecules RhB, with the inset showing a close-up view.
FIG. 12 is an SEM photograph of the PVA gel substrate of comparative example 1.
FIG. 13 is an SEM photograph of RhB-SMIPs-PVA gel substrate of example 3 with the inset partially enlarged.
Detailed Description
The invention will be described more fully hereinafter with reference to the accompanying drawings. Those skilled in the art will be able to implement the invention based on these teachings. Before the present invention is described in detail with reference to the accompanying drawings, it is to be noted that:
the technical solutions and features provided in the present invention in the respective sections including the following description may be combined with each other without conflict.
Moreover, the embodiments of the present invention described in the following description are generally only some embodiments of the present invention, and not all embodiments. Therefore, all other embodiments obtained by a person of ordinary skill in the art based on the embodiments of the present invention without any creative effort shall fall within the protection scope of the present invention.
With respect to terms and units in the present invention. The terms "comprising," "having," and any variations thereof in the description and claims of this invention and the related sections are intended to cover non-exclusive inclusions.
Example 1
0.2g of PVA is weighed and dissolved in 10mL of ultrapure water to obtain a PVA solution with the mass fraction of 2%, then 0.05g of rhodamine B molecularly imprinted polymer (RhB-SMIPs) powder is weighed and added into 1mL of the PVA solution, and the mixture is stirred to be uniformly dispersed to obtain a mixed solution. Then 0.4g of sodium tetraborate is weighed and dissolved in 10mL of ultrapure water to obtain a sodium tetraborate solution. 1mL of mixed solution of RhB-SMIPs and PVA is mixed with 0.4mL of sodium tetraborate solution, and experiments show that the PVA solution and the sodium tetraborate solution in the mass ratio cannot be gelatinized after being mixed.
Example 2
0.4g of PVA is weighed and dissolved in 10mL of ultrapure water to obtain a PVA solution with the mass fraction of 4%, then 0.05g of RhB-SMIPs powder is weighed and added into 1mL of the PVA solution, and the mixture is stirred to be uniformly dispersed to obtain a mixed solution. Then 0.4g of sodium tetraborate is weighed and dissolved in 10mL of ultrapure water to obtain a sodium tetraborate solution. Mixing 1mL of the mixed solution of RhB-SMIPs and PVA with 0.4mL of sodium tetraborate solution, and stirring with a glass rod until gel with good deformable function is formed, thus obtaining the RhB-SMIPs-PVA gel substrate.
Example 3
0.6g of PVA is weighed and dissolved in 10mL of ultrapure water to obtain a PVA solution with the mass fraction of 6%, then 0.05 of RhB-SMIPs powder is weighed and added into 1mL of the PVA solution, and the mixture is stirred to be uniformly dispersed to obtain a mixed solution. Then 0.4g of sodium tetraborate is weighed and dissolved in 10mL of ultrapure water to obtain a sodium tetraborate solution. Mixing 1mL of mixed solution of RhB-SMIPs and PVA with 0.4mL of sodium tetraborate solution, and stirring with a glass rod until gel with good deformable function is formed, thus obtaining the RhB-SMIPs-PVA gel substrate.
Example 4
0.8g of PVA is weighed and dissolved in 10mL of ultrapure water to obtain a PVA solution with the mass fraction of 8%, then 0.05g of RhB-SMIPs powder is weighed and added into 1mL of the PVA solution, and the mixture is stirred to be uniformly dispersed to obtain a mixed solution. Then 0.4g of sodium tetraborate is weighed and dissolved in 10mL of ultrapure water to obtain a sodium tetraborate solution. Mixing 1mL of mixed solution of RhB-SMIPs and PVA with 0.4mL of sodium tetraborate solution, and stirring with a glass rod until gel with good deformable function is formed, thus obtaining the RhB-SMIPs-PVA gel substrate.
Example 5
Weighing 1g of PVA, dissolving the PVA in 10mL of ultrapure water to obtain a PVA solution with the mass fraction of 10%, then weighing 0.05g of RhB-SMIPs powder, adding the powder into 1mL of PVA solution, and stirring to uniformly disperse the powder to obtain a mixed solution. Then 0.4g of sodium tetraborate is weighed and dissolved in 10mL of ultrapure water to obtain a sodium tetraborate solution. Mixing 1mL of mixed solution of RhB-SMIPs and PVA with 0.4mL of sodium tetraborate solution, and stirring with a glass rod until gel with good deformable function is formed, thus obtaining the RhB-SMIPs-PVA gel substrate.
As shown in FIG. 1, it was found that the fluorescence signal was the best when the PVA mass ratio was 6%, when 0.5mg/L, 1mg/L, and 5mg/L of the RhB standard solutions were prepared and dropped on the gel substrates of examples 1 to 5, i.e., the RhB-SMIPs-PVA was subjected to fluorescence detection.
Example 6
0.6g of PVA is weighed and dissolved in 10mL of ultrapure water to obtain a PVA solution with the mass fraction of 6%, then 0.005g of RhB-SMIPs powder is weighed and added into 1mL of the PVA solution, and the mixture is stirred to be uniformly dispersed to obtain a mixed solution. Then 0.4g of sodium tetraborate is weighed and dissolved in 10mL of ultrapure water to obtain a sodium tetraborate solution. Mixing 1mL of mixed solution of RhB-SMIPs and PVA with 0.4mL of sodium tetraborate solution, and stirring with a glass rod until gel with good deformable function is formed, thus obtaining the RhB-SMIPs-PVA gel substrate.
Example 7
0.6g of PVA is weighed and dissolved in 10mL of ultrapure water to obtain a PVA solution with the mass fraction of 6%, then 0.01g of RhB-SMIPs powder is weighed and added into 1mL of the PVA solution, and the mixture is stirred to be uniformly dispersed to obtain a mixed solution. Then 0.4g of sodium tetraborate is weighed and dissolved in 10mL of ultrapure water to obtain a sodium tetraborate solution. Mixing 1mL of mixed solution of RhB-SMIPs and PVA with 0.4mL of sodium tetraborate solution, and stirring with a glass rod until gel with good deformable function is formed, thus obtaining the RhB-SMIPs-PVA gel substrate.
Example 8
0.6g of PVA is weighed and dissolved in 10mL of ultrapure water to obtain a PVA solution with the mass fraction of 6%, then 0.1g of RhB-SMIPs powder is weighed and added into 1mL of the PVA solution, and the mixture is stirred to be uniformly dispersed to obtain a mixed solution. Then 0.4g of sodium tetraborate is weighed and dissolved in 10mL of ultrapure water to obtain a sodium tetraborate solution. Mixing 1mL of mixed solution of RhB-SMIPs and PVA with 0.4mL of sodium tetraborate solution, and stirring with a glass rod until gel with good deformable function is formed, thus obtaining the RhB-SMIPs-PVA gel substrate.
As shown in FIG. 2, it was found that the fluorescence signal was the best when the amount of the RhB-SMIPs powder added was 0.05g, when 0.5mg/L, 1mg/L, and 5mg/L of the RhB standard solutions were prepared, dropped on the RhB-SMIPs-PVA gel substrates of examples 3 and 6 to 8, and subjected to fluorescence detection.
Example 9
0.6g of PVA is weighed and dissolved in 10mL of ultrapure water to obtain a PVA solution with the mass fraction of 6%, then 0.05g of RhB-SMIPs powder is weighed and added into 1mL of the PVA solution, and the mixture is stirred to be uniformly dispersed to obtain a mixed solution. Then 0.4g of sodium tetraborate is weighed and dissolved in 10mL of ultrapure water to obtain a sodium tetraborate solution. Mixing 1mL of mixed solution of RhB-SMIPs and PVA with 0.2mL of sodium tetraborate solution, and stirring with a glass rod until gel with good deformable function is formed, thus obtaining the RhB-SMIPs-PVA gel substrate.
Example 10
0.6g of PVA is weighed and dissolved in 10mL of ultrapure water to obtain a PVA solution with the mass fraction of 6%, then 0.05g of RhB-SMIPs powder is weighed and added into 1mL of the PVA solution, and the mixture is stirred to be uniformly dispersed to obtain a mixed solution. Then 0.4g of sodium tetraborate is weighed and dissolved in 10mL of ultrapure water to obtain a sodium tetraborate solution. Mixing 1mL of mixed solution of RhB-SMIPs and PVA with 0.6mL of sodium tetraborate solution, and stirring with a glass rod until gel with good deformable function is formed, thus obtaining the RhB-SMIPs-PVA gel substrate.
Example 11
0.6g of PVA is weighed and dissolved in 10mL of ultrapure water to obtain a PVA solution with the mass fraction of 6%, then 0.05g of RhB-SMIPs powder is weighed and added into 1mL of the PVA solution, and the mixture is stirred to be uniformly dispersed to obtain a mixed solution. Then 0.4g of sodium tetraborate is weighed and dissolved in 10mL of ultrapure water to obtain a sodium tetraborate solution. Mixing 1mL of mixed solution of RhB-SMIPs and PVA with 0.8mL of sodium tetraborate solution, and stirring with a glass rod until gel with good deformable function is formed, thus obtaining the RhB-SMIPs-PVA gel substrate.
Example 12
0.6g of PVA is weighed and dissolved in 10mL of ultrapure water to obtain a PVA solution with the mass fraction of 6%, then 0.05g of RhB-SMIPs powder is weighed and added into 1mL of the PVA solution, and the mixture is stirred to be uniformly dispersed to obtain a mixed solution. Then 0.4g of sodium tetraborate is weighed and dissolved in 10mL of ultrapure water to obtain a sodium tetraborate solution. And mixing 1mL of mixed solution of RhB-SMIPs and PVA with 1mL of sodium tetraborate solution, and stirring with a glass rod until gel with a good deformable function is formed, thus obtaining the RhB-SMIPs-PVA gel substrate.
0.5mg/L, 1mg/L, and 5mg/L of RhB standard solutions were prepared, and dropped on the RhB-SMIPs-PVA gel substrates of examples 3 and 9 to 12, respectively, to perform fluorescence detection, and the results are shown in FIG. 3. The abscissa in fig. 3 is the volume ratio of the PVA solution to the sodium tetraborate solution, with the corresponding mass ratios of 3:0.4, 3:0.8, 3:1.2, 3:1.6, and 3:2, respectively. As can be seen from FIG. 3, the fluorescence signal is best when the volume ratio of the PVA solution to the sodium tetraborate solution is 10:4 (mass ratio is 3: 0.8).
In order to detect the minimum detection limit of the RhB-SMIPs-PVA gel substrate of the invention on RhB, 0.1-10mg/L concentration gradient RhB solution is prepared, 5uL of the RhB solution is respectively dripped on 0.03g of the RhB-SMIPs-PVA gel substrate of the embodiment 3, and after complete adsorption, fluorescence detection is carried out. As shown in FIG. 4, it can be seen that the RhB-SMIPs-PVA gel substrate can detect RhB in RhB solution with a concentration of 0.1 mg/L. Compared with the RhB with the lowest concentration of 0.1mg/L, which can be detected by the molecular imprinting polymer of the rhodamine B prepared by Zhao morning and the like (Zhao morning, Jiaguanfeng, Lu Wen general, and the like, the preparation of the molecular imprinting polymer of the rhodamine B surface and the fluorescence detection thereof [ J ] food science, 2014,35(20):236 plus 241), the RhB-SMIPs-PVA gel substrate has the advantages that the specificity of the RhB-SMIPs is not influenced, and the detection speed is obviously improved.
To verify the specificity of RhB-SMIPs-PVA gel substrates for RhB adsorption, we selected a dye similar in structure to RhB, namely rhodamine 6G (Rh6G), as a control. Since the molecular structure of Rh6G is very similar to that of RhB, and the molecular structure has strong fluorescence in aqueous solution, the fluorescence wavelength range is also close to that of RhB, and therefore the detection of RhB is often interfered. The specific process is as follows:
the control example of example 3, control example 1, was first conducted, except that RhB-SMIPs were not included in control example 1. Then, the solutions of RhB and Rh6G with concentrations of 10mg/L, 20mg/L, 40mg/L, 50mg/L and 80mg/L were desorbed by the PVA gel substrate of comparative example 1 and the RhB-SMIPs-PVA gel substrate of example 3, respectively, and finally fluorescence detection was performed, and the detection results are shown in FIG. 5 and FIG. 6.
The specificity of RhB-SMIPs can be determined by the enhancement factor α:
α＝IM/IN
in the formula: IM represents the intensity of the fluorescence signal measured with the RhB-SMIPs-PVA gel substrate of example 3, and IN represents the intensity of the fluorescence signal measured with the PVA gel substrate of comparative example 1.
The results of the test after conversion to the enhancer are shown in Table 1.
TABLE 1
As can be seen from the α values of RhB and Rh6G in table 1, the α value of RhB is always greater than that of Rh6G, indicating that the MIP material has a significant effect on the adsorption of RhB in the gel.
Meanwhile, as other red dyes similar to RhB are often added to foods on the market, so that interference is caused on RhB detection, Sunset yellow (Sunset yellow), Sudan I (Sudan I) and Sudan IV (Sudan IV) which are commonly used for food dyeing and have similar colors with RhB and are interfered in the RhB detection process are selected as comparison. FIG. 7 shows the adsorption of RhB, sunset yellow, Sudan I, Sudan IV on the PVA gel substrate of comparative example 1 and the RhB-SMIPs-PVA gel substrate of example 3 at the same concentration of 10 mg/L. It can be seen that in the gel matrix with MIP material, RhB signal is significantly enhanced compared to the other three pigments.
We also evaluated the results by calculating the enhancement factor, which results are shown in table 2.
TABLE 2
As can be seen from Table 2, the enhancement factor of RhB is highest, while the enhancement factors of other pigments are almost equal to 1, which means that the RhB-SMIPs-PVA gel substrate of example 3 has strong interference resistance to RhB.
To verify that the RhB-SMIPs-PVA gel substrate can be well applied to the detection of actual samples, the following tests were performed: 0.5g of chilli powder is respectively weighed and soaked in 0.5mg/L, 1mg/L, 5mg/L and 10mg/L of RhB solution to prepare a standard sample, and the standard sample is dried for standby. Spraying a proper amount of alcohol on a sample to be detected, repeatedly wiping 0.03g of the RhB-SMIPs-PVA gel substrate of the embodiment 3 on the surface of the sample to be detected through dust-free paper, enriching RhB on the sample, and finally placing the sampled substrate on a solid support for fluorescence detection. The test results are shown in FIG. 8, and it can be seen that this method can detect RhB as low as 0.5mg/kg on the actual sample. By comparing the investigation and analysis of the content of rhodamine B in dried chili and chili powder on the market in 2015 by Yishuyan et al (Yishu Yan, HeWei Wei, Jiang Guo, etc.. the investigation and analysis of the content of rhodamine B in dried chili and chili powder [ J ]. China food sanitation journal, 2015, 27(3): 297) 301), it can be found that the minimum detection limit of the RhB-SMIPs-PVA gel substrate in the invention is within the RhB detection content range in the actual samples on the market. Compared with a complicated pretreatment method using an ultra-high performance liquid chromatography-mass spectrometry/mass spectrometry combination instrument, the RhB-SMIPs-PVA gel substrate provided by the invention can realize sampling and detection of samples within a few minutes, and a large amount of time is saved.
The preparation method of RhB-SMIPs of the above example is as follows:
take 1.5g of SiO2Adding the particles and 50ml of anhydrous toluene into an erlenmeyer flask, performing ultrasonic treatment for 20min, and magnetically stirring at room temperature for 1h. 5ml of KH570 (silane coupling agent) was added dropwise to an Erlenmeyer flask, and passed throughAfter the nitrogen gas is exhausted, the mixture is sealed and stirred magnetically for 15 hours. Modifying KH570 with SiO2Washing with anhydrous toluene, centrifuging, washing with anhydrous ethanol for multiple times, and centrifuging to obtain SiO2-KH 570. Another Erlenmeyer flask was added with 10ml acetonitrile, 2mg RhB, 94ul MAA, and magnetically stirred at room temperature under nitrogen for 12 h. Then, 24mgSiO was continuously added to the inside2KH570, 0.875ml EGDMA, 15mg AIBN, and magnetic stirring at 75 ℃ under the protection of nitrogen for 24h to obtain RhB-SMIPs. Washing RhB-SMIPs with methanol, and centrifuging after multiple times of ultrasonic treatment until RhB is washed away. Oven drying the eluted RhB-SMIPs, and grinding into powder.
FIG. 9 is SiO2SEM photograph of KH570, it can be seen that KH570 was surface-modified to cover the surface with organic functional groups of the silane coupling agent. FIG. 10 is an SEM photograph of RhB-SMIPs before elution, and it can be seen that by a series of crosslinking, at SiO2The surface is covered with a layer of organic matter. FIG. 11 is an SEM photograph of RhB-SMIPs after washing away the template molecule RhB, and it can be seen that many cavities are formed in the material, leaving binding sites for RhB.
FIG. 12 is an SEM photograph of the PVA gel substrate of comparative example 1, and it can be seen that the PVA gel substrate has a network structure, which provides a high specific surface area for the adsorption of the substrate. FIG. 13 is an SEM photograph of the RhB-SMIPs-PVA gel substrate of example 3, and it can be seen that RhB-SMIPs are uniformly embedded in the PVA gel and play a crucial role in specific adsorption of RhB.
The contents of the present invention have been explained above. Those skilled in the art will be able to implement the invention based on these teachings. All other embodiments, which can be derived by a person skilled in the art from the above description without inventive step, shall fall within the scope of protection of the present invention.
Claims (9)
1. A composite fluorescent substrate characterized by: the gel comprises a gel and a molecularly imprinted polymer material attached to the pore surface of the gel;
the preparation method comprises the following steps:
(1) obtaining a mixed solution containing a gel precursor and a molecularly imprinted polymer material;
(2) adding a coagulant aid for gelling the gel precursor into gel, and stirring until the gel is formed, thereby obtaining the composite fluorescent substrate;
when in use, the composite fluorescent substrate with the required size is cut, then the cut composite fluorescent substrate is wiped on the surface of a sample for sampling, and solid surface fluorescence detection is carried out after sampling;
the gel is PVA gel;
also comprises sodium tetraborate, wherein the mass ratio of PVA to sodium tetraborate is (1-5) to (0.4-2).
2. The composite fluorescent substrate of claim 1, wherein: the molecularly imprinted polymer is rhodamine B molecularly imprinted polymer.
3. The composite fluorescent substrate of claim 2, wherein: the mass ratio of the rhodamine B molecularly imprinted polymer to the PVA is (0.05-1) to (0.2-1).
4. A method of making a composite fluorescent substrate according to any one of claims 1 to 3, comprising the steps of:
(1) obtaining a mixed solution containing a gel precursor and a molecularly imprinted polymer material;
(2) adding a coagulant aid for gelling the gel precursor into gel, and stirring until the gel is formed, thereby obtaining the composite fluorescent substrate.
5. The method of preparing a composite fluorescent substrate according to claim 4, wherein: the molecularly imprinted polymer is rhodamine B molecularly imprinted polymer.
6. The method of preparing a composite fluorescent substrate according to claim 5, wherein: the step (1) specifically comprises the following steps: firstly, obtaining a PVA solution, and then adding a rhodamine B molecularly imprinted polymer into the PVA solution; the mass fraction of PVA in the PVA solution is 4-10%; the sodium tetraborate is prepared into a solution and then added into the PVA solution.
7. The method of preparing a composite fluorescent substrate according to claim 5, wherein: the preparation of the rhodamine B molecularly imprinted polymer is SiO2Is a carrier, methacrylic acid is a functional monomer, ethylene glycol dimethacrylate is a cross-linking agent, acetonitrile is a pore-foaming agent and 2, 2' -azobisisobutyronitrile is an initiator.
8. The method for detecting the rhodamine B comprises the following steps:
(1) cutting a composite fluorescent substrate, wherein the composite fluorescent substrate is the composite fluorescent substrate in any one of claims 1 to 3 or the composite fluorescent substrate prepared by the preparation method in any one of claims 4 to 7, and the molecularly imprinted polymer is rhodamine B molecularly imprinted polymer;
(2) wiping the surface of the sample to be detected with the cut composite fluorescent substrate;
(3) and carrying out solid surface fluorescence detection on the wiped composite fluorescent substrate to obtain the concentration of rhodamine B on the surface of the sample to be detected.
9. The method for detecting rhodamine B as set forth in claim 8, wherein: the specific process of wiping is as follows: firstly, spraying a solvent on the surface of a sample to be detected, and then wiping the composite fluorescent substrate on the surface of the sample to be detected through dust-free paper.
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1733233A2 (en) * | 2004-03-30 | 2006-12-20 | Whatman, Inc. | Lateral flow format, materials and mehtods |
| CN104165874A (en) * | 2014-07-24 | 2014-11-26 | 江苏大学 | Quantum dot fluorescent aspirin imprinted sensor and its preparation method and use |
| CN104458695A (en) * | 2014-12-02 | 2015-03-25 | 北京理工大学 | Rapid Raman spectrum detection method for molecularly imprinted membrane |
| CN104844762A (en) * | 2015-05-22 | 2015-08-19 | 天津科技大学 | Preparation method for thermo-sensitive type molecularly imprinted polymer with metal frame |
| CN105820585A (en) * | 2016-04-08 | 2016-08-03 | 东北林业大学 | Preparation of mesoporous molecular imprinting fluorescent probe and detection of tetracycline antibiotics by mesoporous molecular imprinting fluorescent probe |
| CN108120707A (en) * | 2018-01-04 | 2018-06-05 | 山西大学 | The preparation of molecular engram fluorescent sensing material and its application in high-throughput detection Rogor |
| CN109370565A (en) * | 2018-11-22 | 2019-02-22 | 江苏大学 | A kind of double emission fluorescent molecularly imprinted polymer nanoparticle and its preparation method and application |
Family Cites Families (20)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7267948B2 (en) * | 1997-11-26 | 2007-09-11 | Ut-Battelle, Llc | SERS diagnostic platforms, methods and systems microarrays, biosensors and biochips |
| NL1016779C2 (en) * | 2000-12-02 | 2002-06-04 | Cornelis Johannes Maria V Rijn | Mold, method for manufacturing precision products with the aid of a mold, as well as precision products, in particular microsieves and membrane filters, manufactured with such a mold. |
| WO2004039940A2 (en) * | 2002-05-14 | 2004-05-13 | Bristol-Myers Squibb Company | Polynucleotide encoding novel human g-protein coupled receptors, and splice variants thereof |
| CN101421035B (en) * | 2006-02-21 | 2015-03-18 | 米普萨卢斯公司 | Improved preparation of molecular imprinted polymers |
| CN100382869C (en) * | 2006-03-13 | 2008-04-23 | 中山大学 | A kind of preparation method of molecularly imprinted solid-phase microextraction coating |
| JP5264421B2 (en) * | 2007-11-22 | 2013-08-14 | 富士フイルム株式会社 | Test substance measuring carrier and method for producing the same |
| CN101581673B (en) * | 2009-06-04 | 2010-11-03 | 江南大学 | Sensor based on iridium composition electroluminescence and molecular imprinting recognition technology |
| CN101696281B (en) * | 2009-10-27 | 2011-07-20 | 中山大学 | Method for preparing molecularly imprinted microgel with fluorescence detection function |
| EP2682438B1 (en) * | 2011-02-28 | 2017-04-05 | FUJIFILM Corporation | Ink composition and image forming method |
| CN103570879A (en) * | 2012-07-26 | 2014-02-12 | 中国科学院沈阳应用生态研究所 | Molecularly-imprinted bonded dyed polystyrene microsphere and preparation method thereof |
| CN103041445A (en) * | 2013-01-21 | 2013-04-17 | 天津工业大学 | Method for preparing molecular imprinting porous gel membrane for tissue engineering |
| WO2014130793A1 (en) * | 2013-02-21 | 2014-08-28 | The Johns Hopkins University | Epitope-polymer platform for detection of bacterial organisms |
| CN103254539B (en) * | 2013-04-23 | 2015-09-16 | 天津工业大学 | A kind of high-intensity high-tenacity protein molecule engram hybrid gel film and preparation method thereof |
| CN104142361B (en) * | 2014-07-31 | 2016-08-17 | 中南民族大学 | A kind of protein molecule engram poly ion liquid membrane electrochemical sensor |
| TR201909273T4 (en) * | 2015-08-20 | 2019-07-22 | Hoffmann La Roche | Particle-based immunoassay using a pegylated analyte specific binding agent. |
| GB2548886B (en) * | 2016-03-31 | 2019-12-25 | Cambridge Display Tech Ltd | Sensor, method of forming a sensor and use thereof |
| CN107556517A (en) * | 2017-09-01 | 2018-01-09 | 天津科技大学 | A kind of preparation method of the Western blotting gel based on cyclodextrin molecular assembly |
| CN107894453B (en) * | 2017-10-25 | 2020-04-28 | 中国农业大学 | Preparation method of molecular imprinting sensor |
| CN109406474B (en) * | 2018-11-14 | 2021-01-29 | 河南工业大学 | Preparation method and application of aggregation-induced emission-molecular imprinting fluorescence sensor for detecting rhodamine B |
| CN109239045B (en) * | 2018-12-11 | 2019-03-26 | 中国科学院烟台海岸带研究所 | A kind of molecular engram ratio fluorescent sensor and preparation and application |
-
2019
- 2019-03-20 CN CN201910213982.5A patent/CN110018142B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1733233A2 (en) * | 2004-03-30 | 2006-12-20 | Whatman, Inc. | Lateral flow format, materials and mehtods |
| CN104165874A (en) * | 2014-07-24 | 2014-11-26 | 江苏大学 | Quantum dot fluorescent aspirin imprinted sensor and its preparation method and use |
| CN104458695A (en) * | 2014-12-02 | 2015-03-25 | 北京理工大学 | Rapid Raman spectrum detection method for molecularly imprinted membrane |
| CN104844762A (en) * | 2015-05-22 | 2015-08-19 | 天津科技大学 | Preparation method for thermo-sensitive type molecularly imprinted polymer with metal frame |
| CN105820585A (en) * | 2016-04-08 | 2016-08-03 | 东北林业大学 | Preparation of mesoporous molecular imprinting fluorescent probe and detection of tetracycline antibiotics by mesoporous molecular imprinting fluorescent probe |
| CN108120707A (en) * | 2018-01-04 | 2018-06-05 | 山西大学 | The preparation of molecular engram fluorescent sensing material and its application in high-throughput detection Rogor |
| CN109370565A (en) * | 2018-11-22 | 2019-02-22 | 江苏大学 | A kind of double emission fluorescent molecularly imprinted polymer nanoparticle and its preparation method and application |
Non-Patent Citations (5)
| Title |
|---|
| Molecular Imprint Membranes Prepared by the Phase Inversion Precipitation Technique;Hong Ying Wang.et;《Langmuir 》;19961231;第4850–4856页 * |
| Molecularly imprinted poly (vinyl alcohol) films for the selective absorption of glycyrrhizinic acid from aqueous solutions;Silvia Patachia.et;《Environmental Engineering and Management Journal》;20091231;第663-668页 * |
| 分子印迹技术及其应用研究进展;柯珍等;《化学研究与应用》;20181231;第30卷(第06期);第865-874页 * |
| 聚合微凝胶胶体晶体水凝胶的研究与应用;陈茂;《中国博士学位论文全文数据库工程科技Ⅰ辑》;20170715;第B014-91页 * |
| 表面分子印迹材料和技术在分离分析中的应用进展;侯会卿等;《色谱》;20161231;第34卷(第12期);第1206-1214页 * |
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