CN109908874B - Novel MoS2QDs @ MIPs molecularly imprinted polymer and preparation method thereof - Google Patents
Novel MoS2QDs @ MIPs molecularly imprinted polymer and preparation method thereof Download PDFInfo
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Abstract
The invention belongs to the technical field of molecular imprinting technology and biological analysis and detection, and relates to a novel MoS2QDs @ MIPs molecularly imprinted polymer and a preparation method thereof. The preparation method of the invention is to prepare the quantum dot MoS firstly2 QDs, and then wrapping at MoS by embedding amikacin in silica shell2 Obtaining a quantum dot compound on the QDs nucleus, and finally eluting amikacin in the quantum dot compound to obtain the molecular imprinting polymer MoS with the spatial imprinting sites of the amikacin2 QDs @ MIPs. The method is simple and easy to implement, the cost is low, and the obtained MoS2The QDs @ MIPs have stable optical performance, high selectivity on the amikacin to be detected, short response time, low detection cost, high sensitivity and high speed when being used for detecting the amikacin, and can be used for content detection of the amikacin.
Description
Technical Field
The invention belongs to the technical field of molecular imprinting technology and biological analysis and detection, relates to a novel molecular imprinting polymer for antibiotic detection and a preparation method thereof, and particularly relates to a novel MoS2QDs @ MIPs molecularly imprinted polymer and a preparation method thereof.
Background
In recent years, along with the development of society, the continuous progress of science and technology and medical technology, antibiotics are widely used, and the problem of antibiotic abuse also appears, and a series of safety problems are caused by drug-resistant bacteria generated by the antibiotic abuse and part of antibiotics remained in organisms, human bodies and environment, so that how to eliminate the pollution and the residue of antibiotics becomes one of the hot spots of research of domestic and foreign scholars. Because the components in biological, human and environmental samples are very complex and the content of antibiotic substances remained in the samples is relatively low, more problems exist in the detection process of the biological, human and environmental samples, such as poor sensitivity, selectivity, accuracy and stability of the detection method.
Since the study of CdSe quantum dots as biomarkers by Alivisatos et al in 1998, the application of semiconductor quantum dots in analysis and detection has been greatly developed. Quantum Dots (QDs) have unique photoelectric properties such as ultraviolet absorptivity, ultrafast optical nonlinear response, photoluminescence, strong photo-oxidation resistance and the like due to the existence of significant size effect, surface effect and quantum effect, and thus are widely applied. However, the single quantum dot fluorescent probe detection is easily interfered by environmental conditions, and has the defects of poor selectivity and limited stability. To solve this problem, research combining quantum dots with other technologies is increasing.
Molecular Imprinting Technique (MIT) is a technique for preparing polymers having a unique selective recognition function for a specific molecule, and the prepared polymers are called Molecular Imprinted Polymers (MIPs). The excellent optical performance of the quantum dots is combined with the high selectivity of the molecular imprinting technology, the defect of poor selectivity of a quantum dot fluorescent probe is just made up by using the extremely high target positioning and enriching capacity of the MIP, and a novel optical sensor based on the molecular imprinting functionalized quantum dots (QDs @ MIP) can be constructed. The molecularly imprinted polymer material can be customized aiming at a target object, realizes the specific recognition of a target molecule, can be compared with a natural biological recognition system (enzyme and substrate), has the characteristics of simple preparation, good stability (acid and alkali resistance, high temperature, high pressure, organic solvent and harsh environment), long service life, easy storage, low manufacturing cost and the like, is widely applied to the aspects of solid phase extraction, chiral separation, biological antibody simulation, catalysis, organic synthesis and the like, and is a simple, convenient and reliable means for solving the problem of high selective recognition of the specific target molecule in complex systems of environment, biology and the like.
Amikacin is an aminoglycoside antibiotic, has an antibacterial spectrum similar to that of gentamicin, is mainly clinically used for various infections caused by gentamicin and kanamycin-resistant gram-negative bacilli such as escherichia coli, proteus vulgaris and pseudomonas aeruginosa, has adverse reactions mainly causing cochlear nerve damage, and has similar toxicity to ears and kidneys to gentamicin. Amikacin, as an antibiotic, is also harmful to the environment, organisms and human bodies in the process of being widely applied. Although there are many methods for detecting amikacin, such as liquid chromatography, colorimetry, molecular imprinting SPR (surface plasmon resonance) nano-sensor method, the liquid chromatography has the defects that if the flow mode of a mobile phase injected into a detector changes, any material which is diffused and kept for separation obviously causes the expansion of a chromatographic peak and the reduction of chromatographic column efficiency; the colorimetric method has the defects of serious interference, higher purity of the detected component and lower sensitivity; although a new means for rapidly detecting antibiotics is provided for the detection method adopting the molecular imprinting SPR nano sensor, the process is more complicated than the high performance liquid chromatography and the colorimetric method.
Currently, more and more researchers have begun to use molecular imprinting for antibiotic detection. However, in the prior art, no method for detecting amikacin by combining the excellent optical performance of quantum dots and the high selectivity of a molecular imprinting technology is reported.
In summary, the main problems of the prior art are: (1) the single quantum dot fluorescent probe detection is easily interfered by environmental conditions, and has the defects of poor selectivity and limited stability; (2) the single molecularly imprinted polymer technology has high selectivity, but often does not have excellent optical performance; (3) the existing amikacin detection method has high cost, long time and poor sensitivity.
Therefore, the research on the method for rapidly and accurately detecting the amikacin by combining the excellent optical performance of the quantum dots and the high selectivity of the molecular imprinting technology has important significance and prospect in the aspects of human health protection, food safety protection, environmental protection and the like.
Disclosure of Invention
In order to overcome the defects of the prior art and solve the problems in the prior art, the invention aims to provide a novel molecularly imprinted polymer MoS2 The preparation method of QDs @ MIPs comprises the steps of firstly preparing quantum dots MoS2 QDs, then encapsulated in MoS by embedding the antibiotic amikacin in a silica shell2 Formation of molecularly imprinted polymers MoS on QDs nuclei2 QDs @ MIPs, the material has good light stability, high selectivity on the substance to be detected, amikacin, short response time and capability of immediately carrying out response detection.
The technical scheme adopted by the invention is as follows: novel molecularly imprinted polymer MoS2 The preparation method of QDs @ MIPs is characterized by comprising the following steps: firstly, preparing quantum dot MoS2 QDs, then encapsulated in MoS by embedding amikacin in silica shell2 Obtaining a quantum dot compound on the QDs nucleus, and finally eluting amikacin in the quantum dot compound to obtain the molecular imprinting polymer MoS with the spatial imprinting sites of the amikacin2 QDs@MIPs。
Further, the molecular imprinting polymer MoS2 The preparation method of QDs @ MIPs is characterized by comprising the following steps:
S1. MoS2 preparation of QDs
Dissolving L-cysteine in deionized water, and performing ultrasonic treatment to obtain a solution A;
mixing Na2MoO4·2H2Dissolving O in deionized water, performing ultrasonic treatment, and adjusting the pH to 5.8-6.5 by using 1mol/L HCl solution to obtain a solution B;
and dropwise adding the solution B into the solution A, performing ultrasonic treatment to obtain a mixed solution, transferring the mixed solution into a polytetrafluoroethylene high-pressure reaction kettle, and reacting at 200 ℃ for 36 h. After the solution is naturally cooled, filtering with filter paper, then filtering with 0.22 μm filter membrane, and finally dialyzing with dialysis bag with cut-off molecular weight of 2000 to obtain quantum dot MoS2 QDs。
S2. MoS2 Preparation of QDs @ MIPs
Quantum dot MoS2 Adding 6mL of QDs into a 100mL beaker, adding 15mL of cyclohexane, 40 mu L of triton X-1002.8-3.6 mL of 3-aminopropyltriethoxysilane, 10-15.0 mg of amikacin, 50-300 mu L of ammonia water and 100 mu L of ethyl orthosilicate under mechanical stirring, sealing, and stirring overnight. Demulsifying the stirred solution with acetone, removing impurities, centrifuging, removing supernatant to obtain precipitate as quantum dot complex, washing the quantum dot complex with ethanol/acetonitrile solution with volume ratio of ethanol to acetonitrile of 4: 1 as eluent, centrifuging at high speed, repeatedly washing the precipitate with ethanol/acetonitrile solution with volume ratio of ethanol to acetonitrile of 4: 1 until amikacin is not detected in supernatant, centrifuging at high speed, and drying the precipitate in oven at 80 deg.C for 12 hr to obtain MoS2 QDs @ MIPs powder.
Further, in step S2, the centrifugation is specifically performed by using a high-speed centrifuge of 2000 revolutions for 2 min.
Further, in step S2, the detecting is specifically performed by using an ultraviolet-visible spectrophotometer.
Further, the operation of step S1 is as follows:
S1. MoS2 preparation of QDs
Dissolving 0.5 g of L-cysteine in 50 mL of deionized water, and carrying out ultrasonic treatment for 10 min to obtain a solution A;
mixing Na2MoO4·2H2Dissolving 0.25 g of O in 25 mL of deionized water, performing ultrasonic treatment for 10 min, and adjusting the pH to 5.8-6.5 by using 1mol/L HCl to obtain a solution B;
and dropwise adding the solution B into the solution A, performing ultrasonic treatment for 10 min to obtain a mixed solution, transferring the mixed solution into a polytetrafluoroethylene high-pressure reaction kettle, and reacting at 200 ℃ for 36 h. After the solution is naturally cooled, filtering with filter paper, filtering with 0.22 μm filter membrane, and dialyzing with dialysis bag with cut-off molecular weight of 2000 to obtain MoS2Quantum dots (MoS)2 QDs）。
The invention also provides a novel molecular imprinting polymer MoS2 QDs @ MIPs, characterized in that: from MoS2 QDs nanoparticles are built within MIPs spheres.
Further, the MoS2 The particle size of the QDs nano-particles is 1-10 nm.
Further, the particle size of the MIPs spheres is 15-50 nm.
Further, the molecular imprinting polymer MoS2 QDs @ MIPs is prepared from the molecular imprinting polymer MoS2 QDs @ MIPs.
Preferably, the molecularly imprinted polymer MoS2 QDs @ MIPs are prepared by the following preparation method:
S1. MoS2 preparation of QDs
Dissolving 0.5 g of L-cysteine in 50 mL of deionized water, and carrying out ultrasonic treatment for 10 min to obtain a solution A;
mixing Na2MoO4·2H2Dissolving 0.25 g of O in 25 mL of deionized water, performing ultrasonic treatment for 10 min, and adjusting the pH to 5.8-6.5 by using 1mol/L HCl to obtain a solution B;
and dropwise adding the solution B into the solution A, performing ultrasonic treatment for 10 min to obtain a mixed solution, transferring the mixed solution into a polytetrafluoroethylene high-pressure reaction kettle, and reacting at 200 ℃ for 36 h. After the solution is naturally cooled, filtering with filter paper, and then filtering with 0.22 μmFiltering with a filter membrane, and dialyzing with a dialysis bag with cut-off molecular weight of 2000 to obtain MoS2Quantum dots (MoS)2 QDs）。
S2. MoS2 Preparation of QDs @ MIPs
Quantum dot MoS2 Adding 6mL of QDs into a 100mL beaker, adding 15mL of cyclohexane, 40 mu L of triton X-1002.8-3.6 mL of 3-aminopropyltriethoxysilane, 10-15.0 mg of amikacin, 50-300 mu L of ammonia water and 100 mu L of ethyl orthosilicate under mechanical stirring, sealing, and stirring overnight. Demulsifying the stirred solution with acetone, removing impurities, centrifuging, removing supernatant to obtain precipitate as quantum dot complex, washing the quantum dot complex with ethanol/acetonitrile solution with volume ratio of ethanol to acetonitrile of 4: 1 as eluent, centrifuging at high speed centrifuge of 2000-2 min, repeatedly washing the precipitate with ethanol/acetonitrile solution with volume ratio of ethanol to acetonitrile of 4: 1 until the supernatant is not detected by UV-visible spectrophotometer to amikacin, centrifuging at high speed centrifuge of 2000-2 min, drying at 80 deg.C for 12 h to obtain MoS2 QDs @ MIPs powder.
The invention also provides a novel molecular imprinting polymer MoS2 The application of QDs @ MIPs in detection of the antibiotic amikacin can be applied to detection of the content of the antibiotic amikacin, such as detection of residual amikacin in samples such as environment and food.
The principle of the invention is as follows:
MoS of the invention2QDs @ MIPs molecularly imprinted polymer is prepared by embedding antibiotics into silica shell and wrapping MoS with the antibiotics2QDs are nucleated to form molecularly imprinted polymers, which have high selectivity.
MoS of the invention2In the synthesis process of QDs @ MIPs, triton is selected as a surfactant, cyclohexane is selected as a continuous phase, ammonia water is selected as a catalyst, 3-aminopropyltriethoxysilane is selected as a functional monomer, tetraethoxysilane is selected as a cross-linking agent, and a template molecule is antibiotic amikacin. In the preparation method of the invention, the quantum dots are dispersed in cyclohexane and triton to form gel, and the gel is prepared byHydrolyzing ethyl orthosilicate, and forming a silicon dioxide layer on the surface of the quantum dot by using ammonia water as a catalyst. In the reaction during polymerization, amino group (-NH) in 3-aminopropyltriethoxysilane2) And the functions (-OH and-NH) of the template molecule2) The groups are bonded by hydrogen bonding. Thus, antibiotics were successfully embedded in silica-coated MoS2In QDs. During elution, the hydrogen bonds in the template molecule and 3-aminopropyltriethoxysilane were broken, leaving antibiotic imprinted sites in the silicon matrix. The size and structure of the imprinting site are consistent with those of antibiotics, so that the antibiotics can be specifically detected.
The invention has the beneficial effects.
1. MoS of the invention2When the QDs @ MIPs is used for detecting antibiotics, the detection cost is low, the detection sensitivity is high, the detection speed is high, and the detection efficiency is high.
2. MoS of the invention2QDs @ MIPs have specific selectivity for analytes, can realize high acceptance of amikacin in several similar antibiotics Streptomycin Sulfate (SS), Kanamycin Sulfate (KS), Tobramycin Sulfate (TS), gentamicin sulfate (QS), ribostamycin sulfate (HS) and Neomycin Sulfate (NS), and can realize selective detection.
3. MoS of the invention2QDs @ MIPs can keep stable optical performance within 18 days, have good stability and are easy to store.
4. MoS of the invention2The preparation method of QDs @ MIPs is simple and feasible and has low cost.
Drawings
FIG. 1(A) is a MoS prepared according to example 1 of the present invention2Transmission electron microscopy of QDs @ MIPs molecularly imprinted polymer;
FIG. 1(B) is a MoS prepared according to comparative example 1 of the present invention2Transmission electron microscopy images of QDs @ NIPs non-molecularly imprinted polymers;
FIG. 1(C) is a MoS prepared according to example 2 of the present invention2Transmission electron microscopy of QDs @ MIPs molecularly imprinted polymer;
FIG. 1(D) is a drawing for illustrating the practice of the present inventionMoS prepared in example 32Transmission electron microscopy of QDs @ MIPs molecularly imprinted polymer;
FIG. 1(E) is a MoS prepared according to example 4 of the present invention2Transmission electron microscopy of QDs @ MIPs molecularly imprinted polymer;
FIG. 1(F) is a MoS prepared according to example 5 of the present invention2Transmission electron microscopy of QDs @ MIPs molecularly imprinted polymer;
FIG. 2(A) is a MoS prepared according to example 1 of the present invention2A fluorescence spectrum diagram of interaction of QDs @ MIPs molecularly imprinted polymer and antibiotics;
FIG. 2(B) is a MoS prepared according to example 1 of the present invention2A linear fit graph of the interaction of QDs @ MIPs molecularly imprinted polymers and antibiotics;
FIG. 3(A) is a MoS prepared according to comparative example 1 of the present invention2Fluorescence spectra of interaction of QDs @ NIPs non-molecularly imprinted polymers and antibiotics;
FIG. 3(B) is a MoS prepared according to comparative example 1 of the present invention2A linear fit plot of QDs @ NIPs non-molecularly imprinted polymer interactions with antibiotics;
FIG. 4 is a MoS prepared according to example 1 of the present invention2Fluorescence stability diagram of QDs @ MIPs molecularly imprinted polymer at room temperature;
FIG. 5 is a MoS prepared according to example 1 of the present invention2Plots of the selective response of QDs @ MIPs molecularly imprinted polymers to several similar antibiotics.
Detailed Description
The technical solution of the present invention will be described in detail with reference to the accompanying drawings, but the present invention is not limited thereto.
Example 1:
MoS2preparation of QDs @ MIPs
S1. MoS2 Preparation of QDs
Dissolving 0.5 g of L-cysteine in 50 mL of deionized water, and carrying out ultrasonic treatment for 10 min to obtain a solution A.
Mixing Na2MoO4·2H2Dissolving 0.25 g of O in 25 mL of deionized water, carrying out ultrasonic treatment for 10 min, and adjusting the pH to 5.8-6.5 by using 1mol/L HCl to obtain a solution B.
Mixing the solutionAnd B, dropwise adding the solution A into the solution B, performing ultrasonic treatment for 10 min to obtain a mixed solution, transferring the mixed solution into a polytetrafluoroethylene high-pressure reaction kettle, and reacting at 200 ℃ for 36 h. After the solution is naturally cooled, filtering with filter paper, filtering with 0.22 μm filter membrane, and dialyzing with dialysis bag with cut-off molecular weight of 2000 to obtain MoS2Quantum dots (MoS)2 QDs）。
S2. MoS2 Preparation of QDs @ MIPs
Quantum dot MoS2 QDs 6mL was added to a 100mL beaker, cyclohexane 15mL, Triton X-1002.8 mL, 3-aminopropyltriethoxysilane 40. mu.L, amikacin 12 mg, ammonia 200. mu.L, and ethyl orthosilicate 100. mu.L were added with mechanical stirring, sealed, and stirred overnight. Demulsifying the stirred solution with acetone, removing impurities, centrifuging at 2000-2 min with a high-speed centrifuge, discarding supernatant to obtain quantum dot complex, washing the quantum dot complex with ethanol/acetonitrile solution with volume ratio of ethanol to acetonitrile of 4: 1 as eluent, centrifuging at 2000-2 min with a high-speed centrifuge, repeatedly washing the precipitate with ethanol/acetonitrile solution with volume ratio of ethanol to acetonitrile of 4: 1 until the supernatant is not detected by UV-visible spectrophotometer, centrifuging at 2000-2 min with a high-speed centrifuge, drying at 80 deg.C for 12 h to obtain MoS2 QDs @ MIPs molecularly imprinted polymer powder.
MoS prepared from example 12A transmission electron micrograph of the QDs @ MIPs molecularly imprinted polymer is shown in FIG. 1 (A).
Comparative example 1
Non-molecularly imprinted polymers (MoS)2QDs @ NIPs):
prepared using the same method as example 1, but without the addition of the template molecule amikacin.
Dissolving 0.5 g of L-cysteine in 50 mL of deionized water, and carrying out ultrasonic treatment for 10 min to obtain a solution A.
Mixing Na2MoO4·2H2Dissolving 0.25 g of O in 25 mL of deionized water, carrying out ultrasonic treatment for 10 min, and adjusting the pH to 5.8-6.5 by using 1mol/L HCl to obtain a solution B.
And dropwise adding the solution B into the solution A, performing ultrasonic treatment for 10 min to obtain a mixed solution, transferring the mixed solution into a polytetrafluoroethylene high-pressure reaction kettle, and reacting at 200 ℃ for 36 h. After the solution is naturally cooled, filtering with filter paper, filtering with 0.22 μm filter membrane, and dialyzing with dialysis bag with cut-off molecular weight of 2000 to obtain MoS2Quantum dots (MoS)2 QDs）。
S2. MoS2 Preparation of QDs @ NIPs
Quantum dot MoS2 QDs 6mL was added to a 100mL beaker, and under mechanical stirring, cyclohexane 15mL, Triton X-1002.8 mL, 3-aminopropyltriethoxysilane 40. mu.L, ammonia 200. mu.L, and tetraethoxysilane 100. mu.L were added, sealed, and stirred overnight. Demulsifying the stirred solution with acetone, removing impurities, centrifuging at 2000-2 min with a high-speed centrifuge, discarding supernatant to obtain quantum dot complex, washing the quantum dot complex with ethanol/acetonitrile solution with volume ratio of ethanol to acetonitrile of 4: 1 as eluent, centrifuging at 2000-2 min with a high-speed centrifuge, repeatedly washing the precipitate with ethanol/acetonitrile solution with volume ratio of ethanol to acetonitrile of 4: 1 until the supernatant is not detected by UV-visible spectrophotometer, centrifuging at 2000-2 min with a high-speed centrifuge, drying at 80 deg.C for 12 h to obtain MoS2 QDs @ NIPs non-molecularly imprinted polymer powders.
MoS prepared from comparative example 12A transmission electron micrograph of non-molecularly imprinted polymers of QDs @ NIPs is shown in FIG. 1 (B).
Example 2:
MoS2preparation of QDs @ MIPs
S1. MoS2 Preparation of QDs
Dissolving 0.5 g of L-cysteine in 50 mL of deionized water, and carrying out ultrasonic treatment for 10 min to obtain a solution A.
Mixing Na2MoO4·2H2Dissolving 0.25 g of O in 25 mL of deionized water, carrying out ultrasonic treatment for 10 min, and adjusting the pH to 5.8-6.5 by using 1mol/L HCl to obtain a solution B.
Dropwise adding the solution B into the solution A, and performing ultrasonic treatment for 10 min to obtain a mixtureAnd transferring the mixed solution into a polytetrafluoroethylene high-pressure reaction kettle, and reacting at 200 ℃ for 36 h. After the solution is naturally cooled, filtering with filter paper, filtering with 0.22 μm filter membrane, and dialyzing with dialysis bag with cut-off molecular weight of 2000 to obtain MoS2Quantum dots (MoS)2 QDs）。
S2. MoS2 Preparation of QDs @ MIPs
Quantum dot MoS2 QDs 6mL was added to a 100mL beaker, cyclohexane 15mL, Triton X-1003.6 mL, 3-aminopropyltriethoxysilane 40. mu.L, amikacin 10 mg, ammonia 200. mu.L, and ethyl orthosilicate 100. mu.L were added with mechanical stirring, sealed, and stirred overnight. Demulsifying the stirred solution with acetone, removing impurities, centrifuging at 2000-2 min with a high-speed centrifuge, discarding supernatant to obtain quantum dot complex, washing the quantum dot complex with ethanol/acetonitrile solution with volume ratio of ethanol to acetonitrile of 4: 1 as eluent, centrifuging at 2000-2 min with a high-speed centrifuge, repeatedly washing the precipitate with ethanol/acetonitrile solution with volume ratio of ethanol to acetonitrile of 4: 1 until the supernatant is not detected by UV-visible spectrophotometer, centrifuging at 2000-2 min with a high-speed centrifuge, drying at 80 deg.C for 12 h to obtain MoS2 QDs @ MIPs molecularly imprinted polymer powder.
MoS prepared from example 22A transmission electron micrograph of the QDs @ MIPs molecularly imprinted polymer is shown in FIG. 1 (C).
Example 3:
MoS2preparation of QDs @ MIPs
S1. MoS2 Preparation of QDs
Dissolving 0.5 g of L-cysteine in 50 mL of deionized water, and carrying out ultrasonic treatment for 10 min to obtain a solution A.
Mixing Na2MoO4·2H2Dissolving 0.25 g of O in 25 mL of deionized water, carrying out ultrasonic treatment for 10 min, and adjusting the pH to 5.8-6.5 by using 1mol/L HCl to obtain a solution B.
Dropwise adding the solution B into the solution A, performing ultrasonic treatment for 10 min to obtain a mixed solution, and transferring the mixed solution to polytetrafluoroethylene high-pressure reactorReacting for 36 h at 200 ℃ in a kettle. After the solution is naturally cooled, filtering with filter paper, filtering with 0.22 μm filter membrane, and dialyzing with dialysis bag with cut-off molecular weight of 2000 to obtain MoS2Quantum dots (MoS)2 QDs）。
S2. MoS2 Preparation of QDs @ MIPs
Quantum dot MoS2 QDs 6mL was added to a 100mL beaker, cyclohexane 15mL, Triton X-1003.0 mL, 3-aminopropyltriethoxysilane 40. mu.L, amikacin 15.0 mg, ammonia 200. mu.L, and ethyl orthosilicate 100. mu.L were added with mechanical stirring, sealed, and stirred overnight. Demulsifying the stirred solution with acetone, removing impurities, centrifuging at 2000-2 min with a high-speed centrifuge, discarding supernatant to obtain quantum dot complex, washing the quantum dot complex with ethanol/acetonitrile solution with volume ratio of ethanol to acetonitrile of 4: 1 as eluent, centrifuging at 2000-2 min with a high-speed centrifuge, repeatedly washing the precipitate with ethanol/acetonitrile solution with volume ratio of ethanol to acetonitrile of 4: 1 until the supernatant is not detected by UV-visible spectrophotometer, centrifuging at 2000-2 min with a high-speed centrifuge, drying at 80 deg.C for 12 h to obtain MoS2 QDs @ MIPs molecularly imprinted polymer powder.
MoS prepared from example 32A transmission electron micrograph of the QDs @ MIPs molecularly imprinted polymer is shown in FIG. 1 (D).
Example 4:
MoS2preparation of QDs @ MIPs
S1. MoS2 Preparation of QDs
Dissolving 0.5 g of L-cysteine in 50 mL of deionized water, and carrying out ultrasonic treatment for 10 min to obtain a solution A.
Mixing Na2MoO4·2H2Dissolving 0.25 g of O in 25 mL of deionized water, carrying out ultrasonic treatment for 10 min, and adjusting the pH to 5.8-6.5 by using 1mol/L HCl to obtain a solution B.
And dropwise adding the solution B into the solution A, performing ultrasonic treatment for 10 min to obtain a mixed solution, transferring the mixed solution into a polytetrafluoroethylene high-pressure reaction kettle, and reacting at 200 ℃ for 36 h. Waiting for the solution to be naturalAfter cooling, filtering with filter paper, filtering with 0.22 μm filter membrane, and dialyzing with dialysis bag with cut-off molecular weight of 2000 to obtain MoS2Quantum dots (MoS)2 QDs）。
S2. MoS2 Preparation of QDs @ MIPs
Quantum dot MoS2 QDs 6mL was added to a 100mL beaker, cyclohexane 15mL, Triton X-1002.8 mL, 3-aminopropyltriethoxysilane 40. mu.L, amikacin 12.0 mg, ammonia 50. mu.L, and ethyl orthosilicate 100. mu.L were added with mechanical stirring, sealed, and stirred overnight. Demulsifying the stirred solution with acetone, removing impurities, centrifuging at 2000-2 min with a high-speed centrifuge, discarding supernatant to obtain quantum dot complex, washing the quantum dot complex with ethanol/acetonitrile solution with volume ratio of ethanol to acetonitrile of 4: 1 as eluent, centrifuging at 2000-2 min with a high-speed centrifuge, repeatedly washing the precipitate with ethanol/acetonitrile solution with volume ratio of ethanol to acetonitrile of 4: 1 until the supernatant is not detected by UV-visible spectrophotometer, centrifuging at 2000-2 min with a high-speed centrifuge, drying at 80 deg.C for 12 h to obtain MoS2 QDs @ MIPs molecularly imprinted polymer powder.
MoS prepared from example 42A transmission electron micrograph of the QDs @ MIPs molecularly imprinted polymer is shown in FIG. 1 (E).
Example 5:
MoS2preparation of QDs @ MIPs
S1. MoS2 Preparation of QDs
Dissolving 0.5 g of L-cysteine in 50 mL of deionized water, and carrying out ultrasonic treatment for 10 min to obtain a solution A.
Mixing Na2MoO4·2H2Dissolving 0.25 g of O in 25 mL of deionized water, carrying out ultrasonic treatment for 10 min, and adjusting the pH to 5.8-6.5 by using 1mol/L HCl to obtain a solution B.
And dropwise adding the solution B into the solution A, performing ultrasonic treatment for 10 min to obtain a mixed solution, transferring the mixed solution into a polytetrafluoroethylene high-pressure reaction kettle, and reacting at 200 ℃ for 36 h. After the solution is naturally cooled, filtering with filter paper, and then filtering with 0.22 μm filter membraneFiltering, and dialyzing with dialysis bag with cut-off molecular weight of 2000 to obtain MoS2Quantum dots (MoS)2 QDs）。
S2. MoS2 Preparation of QDs @ MIPs
Quantum dot MoS2 QDs 6mL was added to a 100mL beaker, cyclohexane 15mL, Triton X-1002.8 mL, 3-aminopropyltriethoxysilane 40. mu.L, amikacin 12.0 mg, ammonia 100. mu.L, and ethyl orthosilicate 100. mu.L were added with mechanical stirring, sealed, and stirred overnight. Demulsifying the stirred solution with acetone, removing impurities, centrifuging at 2000-2 min with a high-speed centrifuge, discarding supernatant to obtain quantum dot complex, washing the quantum dot complex with ethanol/acetonitrile solution with volume ratio of ethanol to acetonitrile of 4: 1 as eluent, centrifuging at 2000-2 min with a high-speed centrifuge, repeatedly washing the precipitate with ethanol/acetonitrile solution with volume ratio of ethanol to acetonitrile of 4: 1 until the supernatant is not detected by UV-visible spectrophotometer, centrifuging at 2000-2 min with a high-speed centrifuge, drying at 80 deg.C for 12 h to obtain MoS2 QDs @ MIPs molecularly imprinted polymer powder.
MoS prepared from example 52A transmission electron micrograph of the QDs @ MIPs molecularly imprinted polymer is shown in FIG. 1 (F).
Example 6:
(1) MoS2QDs @ MIPs performance test
To 10 mL of phosphate buffer (pH = 7.0) was added MoS obtained in example 12QDs @ MIPs molecularly imprinted polymer 2.0 mg, ultrasonically vibrated for 20 minutes. 1mL of the above solution was added to 16 10 mL cuvettes, and 1mL of a series of standard solutions of amikacin with different concentrations were added, wherein the concentrations of the standard solutions of amikacin were 0.5nmol/L, 1nmol/L, 2nmol/L, 5nmol/L, 10nmol/L, 15nmol/L, 20nmol/L, 30nmol/L, 40nmol/L, 50nmol/L, 60nmol/L, 80nmol/L, 100nmol/L, 200nmol/L, 500nmol/L, and 1000nmol/L, respectively. And (4) uniformly mixing. Adjusting the excitation wavelength to 350nm with fluorescence spectrophotometer, selecting the spectral range at 390nm and 560nm, and measuring the above series of solutionsFluorescence emission spectrum of the liquid. Observing the fluorescence spectra of corresponding systems with different concentrations of amikacin, and observing whether the fluorescence intensity of the amikacin generates quenching or not, thereby determining the MoS2QDs @ MIPs have selective performance. MoS2Fluorescence spectra of the interaction of QDs @ MIPs with antibiotics, see fig. 2 (a); a linear fit of the fluorescence intensity ratio as a function of amikacin concentration was also plotted, see FIG. 2 (B).
As can be seen from FIGS. 2(A) and 2(B), under the same detection conditions, MoS2The QDs @ MIPs molecularly imprinted polymer has obvious fluorescent response to template molecule amikacin, and system fluorescence generates quenching, namely MoS2QDs @ MIPs have a specific recognition effect on the template molecule amikacin. When the concentration of amikacin is 0.5-1000nmol/L, the fluorescence intensity ratio and the concentration of amikacin are in a linear relation, and the linear equation is as follows: y =1.208+0.0150x, R2=0.9948, and from this, it was found that the fluorescence intensity ratio and amikacin have a good linear relationship. Tested, MoS2The detection limit of the QDs @ MIPs molecularly imprinted polymer on amikacin is 0.23 nmol/L.
(2)MoS2Performance assays for QDs @ NIPs
To 10 mL of phosphate buffer (pH = 7.0) was added MoS obtained in comparative example 12QDs @ NIPs non-molecularly imprinted polymer 2.0 mg, ultrasonically vibrated for 20 minutes. 1mL of the solution is added into 9 10 mL colorimetric tubes respectively, and a series of amikacin standard solutions with different concentrations, namely 1mL, are added, wherein the concentrations of the amikacin standard solutions are 0.5nmol/L, 1nmol/L, 2nmol/L, 4nmol/L, 6nmol/L, 8nmol/L, 10nmol/L, 12nmol/L and 15nmol/L in sequence. And (4) uniformly mixing. And (3) adjusting the excitation wavelength to 350nm by using a fluorescence spectrophotometer, selecting the spectral range between 390nm and 560nm, and measuring the fluorescence emission spectrum of the series of solutions. Observing the fluorescence spectra of corresponding systems with different concentrations of amikacin, and observing whether the fluorescence intensity of the amikacin generates quenching or not, thereby determining the MoS2QDs @ NIPs has selective properties. MoS2Fluorescence spectra of the interaction of QDs @ NIPs with antibiotics, see FIG. 3 (A); a linear fit of the fluorescence intensity ratio as a function of amikacin concentration was also plotted, see FIG. 3 (B).
As can be seen from FIG. 3(A), the fluorescence intensity of the system is almost constant, and from FIGS. 3(A) and 3(B), under the same detection conditions, MoS2The QDs @ NIPs non-molecularly imprinted polymer has no obvious fluorescent response to the template molecule amikacin, namely no quenching is generated on the system fluorescence.
Example 7:
MoS2optical stability of QDs @ MIPs
To test MoS2Optical stability of QDs @ MIPs Using the MoS obtained in example 12QDs @ MIPs molecularly imprinted polymer, measuring the fluorescence spectrum of the molecularly imprinted polymer continuously for 30 days at room temperature, wherein the excitation wavelength is 350nm, and drawing a fluorescence stability graph. MoS at room temperature2Fluorescence stability plots of QDs @ MIPs, see FIG. 4. As can be seen from FIG. 4, MoS at room temperature2The fluorescence intensity of QDs @ MIPs gradually decreased after 18 days, with the spectral width and emission wavelength remaining unchanged. Thus, the MoS obtained was found to be2The QDs @ MIPs molecularly imprinted polymer can keep relatively stable optical performance within 18 days, and the phenomenon also indicates that MoS2QDs @ MIPs have good stability.
Example 8:
MoS2selective detection of QDs @ MIPs
To test MoS2Selectivity of QDs @ MIPs with MoS2QDs @ NIPs, MoS was performed using several antibiotic analogs Streptomycin Sulfate (SS), Kanamycin Sulfate (KS), Tobramycin Sulfate (TS), gentamicin sulfate (QS), ribostamycin sulfate (HS), and Neomycin Sulfate (NS)2QDs @ MIPs and MoS2Selectivity test for QDs @ NIPs for amikacin. The resulting MoS2QDs @ MIPs and MoS2The selective response profile of QDs @ NIPs to several of the above antibiotics is shown in figure 5.
As can be seen from FIG. 5, MoS2The acceptance of QDs @ MIPs for Amikacin (AS) is higher than that for Streptomycin Sulfate (SS), Kanamycin Sulfate (KS), Tobramycin Sulfate (TS), gentamicin sulfate (QS), ribostamycin sulfate (HS) and Neomycin Sulfate (NS), i.e., MoS2QDs @ MIPs have specific recognition only for the template molecule amikacinOther actions are described. And MoS2The fluorescence response of QDs @ NIPs to the antibiotics Amikacin (AS), Streptomycin Sulfate (SS), Kanamycin Sulfate (KS), Tobramycin Sulfate (TS), gentamicin sulfate (QS), ribostamycin sulfate (HS), and Neomycin Sulfate (NS) was nearly identical. These results show that MoS2The recognition mechanism of QDs @ MIPs is based on the interaction of the shape, size and function of the template. In MoS2In QDs @ NIPs, no specific recognition site is formed because no template is added, and thus MoS2The adsorption capacity of QDs @ NIPs to template molecules is far lower than that of MoS2QDs @ MIPs, and no significant difference in quenching ability from other similar antibiotics.
Example 9:
sample analysis
Using the MoS obtained in example 12QDs @ MIPs to detect amikacin in a sample.
A river water sample (3 portions) was boiled and filtered 3 times through a 0.22 μm filter membrane. Using the MoS obtained in example 12QDs @ MIPs as per MoS in example 62The linear equation in the performance test of QDs @ MIPs calculates the concentration of amikacin in the system, calculates the recovery rate according to the additive amount and the measured value, and respectively measures the recovery rate of amikacin in 3 samples, and the result is shown in Table 1.
TABLE 1 determination of Amikacin recovery in samples
As can be seen from Table 1, MoS2The recovery rate of the QDs @ MIPs to the amikacin is 98.8% -103.5%, and the RSD values are all lower than 5.78%, which shows that the method is high in accuracy and good in precision.
The above description is only for the preferred embodiment of the present invention, and is not intended to limit the scope of the present invention. Any technical solution that can be implemented in the scope of the claims covered by the present application, or any technical solution that can be made by the technical personnel skilled in the art by using the contents of the method disclosed in the above, is covered by the scope of the present invention.
Claims (6)
1. Molecularly imprinted polymer MoS2QDs @ MIPs, characterized in that: the molecular imprinting polymer MoS2 The preparation method of QDs @ MIPs specifically comprises the following steps:
S1. MoS2 preparation of QDs
Dissolving L-cysteine in deionized water, and performing ultrasonic treatment to obtain a solution A;
mixing Na2MoO4·2H2Dissolving O in deionized water, performing ultrasonic treatment, and adjusting the pH to 5.8-6.5 by using 1mol/L HCl solution to obtain a solution B;
dropwise adding the solution B into the solution A, performing ultrasonic treatment to obtain a mixed solution, transferring the mixed solution into a polytetrafluoroethylene high-pressure reaction kettle, and reacting at 200 ℃ for 36 h;
after the solution is naturally cooled, filtering with filter paper, then filtering with 0.22 μm filter membrane, and finally dialyzing with dialysis bag with cut-off molecular weight of 2000 to obtain quantum dot MoS2 QDs；
S2. MoS2 Preparation of QDs @ MIPs
Quantum dot MoS2 Adding 6mL of QDs into a 100mL beaker, adding 15mL of cyclohexane, 40 mu L of triton X-1002.8-3.6 mL of 3-aminopropyltriethoxysilane, 10-15.0 mg of amikacin, 50-300 mu L of ammonia water and 100 mu L of ethyl orthosilicate under mechanical stirring, sealing, and stirring overnight;
demulsifying the stirred solution with acetone, removing impurities, centrifuging, removing supernatant to obtain precipitate as quantum dot complex, washing the quantum dot complex with ethanol/acetonitrile solution with volume ratio of ethanol to acetonitrile of 4: 1 as eluent, centrifuging at high speed, repeatedly washing the precipitate with ethanol/acetonitrile solution with volume ratio of ethanol to acetonitrile of 4: 1 until amikacin is not detected in supernatant, centrifuging at high speed, and drying the precipitate in oven at 80 deg.C for 12 hr to obtain MoS2 QDs @ MIPs powder;
the MoS2The QDs @ MIPs molecularly imprinted polymer is used for detecting amikacin with the amikacin concentration ofWhen the concentration is 0.5-1000nmol/L, a fluorescence spectrophotometer is adopted for detection, the excitation wavelength is 350nm, the spectral range is 390nm-560nm, and the detection limit is 0.23 nmol/L.
2. A molecularly imprinted polymer MoS according to claim 12QDs @ MIPs, characterized in that: the operation of step S1 is as follows:
S1. MoS2 preparation of QDs
Dissolving 0.5 g of L-cysteine in 50 mL of deionized water, and carrying out ultrasonic treatment for 10 min to obtain a solution A;
mixing Na2MoO4·2H2Dissolving 0.25 g of O in 25 mL of deionized water, performing ultrasonic treatment for 10 min, and adjusting the pH to 5.8-6.5 by using 1mol/L HCl to obtain a solution B;
dropwise adding the solution B into the solution A, performing ultrasonic treatment for 10 min to obtain a mixed solution, transferring the mixed solution into a polytetrafluoroethylene high-pressure reaction kettle, and reacting at 200 ℃ for 36 h;
after the solution is naturally cooled, filtering with filter paper, then filtering with 0.22 μm filter membrane, and finally dialyzing with dialysis bag with cut-off molecular weight of 2000 to obtain quantum dot MoS2 QDs。
3. A molecularly imprinted polymer MoS according to claim 12QDs @ MIPs, characterized in that: in step S2, the centrifugation is specifically performed by using a high-speed centrifuge for 2 min at 2000 rpm, and the supernatant detection is specifically performed by using an ultraviolet-visible spectrophotometer.
4. A molecularly imprinted polymer MoS according to claim 12QDs @ MIPs, characterized in that: from MoS2 QDs nanoparticles are built within MIPs spheres.
5. A molecularly imprinted polymer MoS according to claim 42QDs @ MIPs, characterized in that: the MoS2 The particle size of the QDs nano-particles is 1-10 nm.
6. A molecularly imprinted polymer MoS according to claim 42QDs @ MIPs, characterized in that: the particle size of the MIPs balls is 15-50 nm.
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