CN111072657A - Fluorescent probe for detecting activity of acetylcholinesterase and synthetic method and application thereof - Google Patents
Fluorescent probe for detecting activity of acetylcholinesterase and synthetic method and application thereof Download PDFInfo
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- C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
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Abstract
The invention discloses a fluorescent probe for detecting the activity of acetylcholinesterase, a synthetic method and application thereof, wherein the invention is based on MnO2The AChE activity detection is realized through the electrostatic interaction between the nanosheets and the cationic derivatives (PBI-DM) of the perylene tetracarboxylic diimide and the fluorescence spectrum characteristics of the PBI-DM in the aggregation and dispersion processes. The invention discloses a synthesized fluorescent probe PBI-DM @ MnO2Detection of AChE by NS has a value of 0.24mLow detection limit of U/mL, high selectivity and PBI-DM @ MnO2NS can be used for screening AChE inhibitors and has great potential in the field of clinical diagnosis.
Description
Technical Field
The invention belongs to the technical field of acetylcholinesterase activity detection, and relates to a method strategy for detecting the activity of acetylcholinesterase (AChE). More particularly, the present invention relates not only to a MnO based2The novel method for detecting the AChE activity through the electrostatic interaction between the nanosheets and the cationic derivative (PBI-DM) of the perylene tetracarboxylic diimide and the fluorescence spectrum characteristics of the PBI-DM in the aggregation and dispersion process further relates to a synthetic method of the fluorescent probe and application of the fluorescent probe in screening of the AChE inhibitor.
Background
In addition, more and more studies indicate that oxidative stress in depression is a major cause of dysfunction of neuronal metabolism in the brain, whereas AChE is a key hydrolase in the cholinergic system that hydrolyzes Acetylcholine (ATCH) to produce thiocholine (TCh), and abnormal fluctuations in AChE directly affect ATCH metabolism, thereby disrupting neurotransmitters in the brain, which inevitably affect mood and behavior, and thus may induce the occurrence of depression.
Nowadays, the detection methods of AChE activity mainly include Ellman test, fluorescence assay method and electrochemical assay. Although the Ellman test is widely used, numerous analytical studies have shown that there are certain drawbacks and limitations in the detection of AChE activity due to its false positive effect and poor accuracy. In addition, the conventional assay methods have lower sensitivity compared to luminescence assays, and therefore, luminescence sensing-based fluorescence assays are one of the most attractive strategies for determining AChE activity due to their ease of development, high sensitivity, good selectivity, and ease of operation.
In conclusion, the technical personnel in the field need to solve the problem of how to provide a fluorescent probe for detecting the activity of acetylcholinesterase with high sensitivity and high selectivity and a synthetic method thereof.
Disclosure of Invention
In view of the above, the present invention provides a MnO-based method for solving the problems in the prior art2The electrostatic interaction between the nanosheets and the cationic derivative of perylene tetracarboxylic diimide (PBI-DM), and the novel method for detecting the AChE activity of the PBI-DM through the fluorescence spectrum characteristics in the aggregation and dispersion process, and the MnO loaded with the PBI-DM are disclosed2Nanosheet (PBI-DM @ MnO)2NS) fluorescent probe.
In order to achieve the purpose, the technical scheme of the invention is as follows:
a method for synthesizing a fluorescent probe for detecting acetylcholinesterase activity comprises the following steps:
the method comprises the following steps: adding PBS buffer solution into a colorimetric tube as a solvent;
step two: adding a DMSO solution of PBI-DM and MnO to the solvent in the step one2Obtaining PBI-DM @ MnO from nanosheet mother liquor through electrostatic adsorption2NS。
The specific preparation scheme is as follows:
the maximum concentration of PBI-DM in PBS buffer (pH 7.4) was 6 × 10 as measured by fluorescence spectroscopy and ultraviolet absorption spectroscopy of PBI-6mol·L-1，MnO2The optimal concentration of nanoplatelets capable of quenching PBI-DM was 7.65 μ g/mL. Thus, 2mL of PBS buffer (pH 7.4) was added as a solvent to a 5mL colorimetric tube, and 2.4 μ L of PBI-DM in DMSO (5 × 10) was added thereto-3mol·L-1) Then, 5.5. mu.L of MnO was added2Uniformly mixing the nanosheet mother liquor (2.78mg/mL) to obtain PBI-DM @ MnO2NS。
Preferably, the concentration of the PBI-DM is 6 x 10-6mol·L-1，MnO2The concentration of the nano-sheets is 7.65 mu g/mL.
In order to achieve the above object, the PBI-DM used in the present invention is prepared by the following method:
the structural formula of the PBI-DM is as follows:
the synthetic route of the PBI-DM is as follows:
the synthesis method of the cationic derivative PBI-DM of perylene tetracarboxylic diimide comprises the following steps:
(1) DMF is taken as a reaction medium, perylene tetracarboxylic anhydride and N, N-dimethylethylamine are taken as substrates, zinc acetate is taken as a catalyst, and the mixture is heated and refluxed at 120 ℃ for reaction overnight;
(2) adding secondary water into the reaction solution obtained in the step (1) for washing, and drying to obtain a reaction product I;
(3) adding a toluene solvent into the reaction product I obtained in the step (2), adding methyl iodide serving as a reaction substrate, and heating and refluxing for 3 hours to obtain a reaction product II;
(4) and (4) adding ether into the reaction product II obtained in the step (3) to filter and wash the precipitate, and performing suction filtration and vacuum drying to obtain a brownish red product and obtain a target pure product PBI-DM.
Preferably, the molar ratio of the perylene tetracarboxylic anhydride to the N, N-dimethylethylamine is 1: 10.
The synthesis method of the cationic derivative PBI-DM of perylene tetracarboxylic diimide disclosed above is not only simple to operate, but also convenient and fast to purify, and is suitable for popularization and application in the market.
In addition, the inventor carries out characterization through means such as nuclear magnetic resonance hydrogen spectrum, carbon spectrum and the like, and shows that the synthesis of PBI-DM is successful, and the specific reference is made to the attached figures 1 and 2 in the specification.
In addition, it is to be noted that MnO used in the present invention is for achieving the above object2The nano-sheet mother liquor adopts the following preparation method:
(1) preparation of a catalyst composition comprisingHas a concentration of 0.6 mol.L-1Tetramethylammonium hydroxide and 3% H2O220mL of mixed aqueous solution;
(2) dropwise adding the reaction solution obtained in the step (1) into 15s until the concentration of 10mL is 0.3 mol.L-1MnCl of2In the solution, the solution turns dark brown, and is stirred vigorously at room temperature overnight;
(3) centrifuging the reaction product crude product obtained in the step (2) at 2000rpm for 10min, washing with a large amount of secondary water and methanol, and drying at 60 ℃ to obtain a target pure product MnO2Nanosheets;
(4) dispersing 0.01g of the reaction product obtained in the step (3) into 40mL of secondary water, performing ultrasonic treatment for 10 hours, centrifuging at 2000rpm, and leaving supernatant to obtain MnO with the concentration of 2.78mg/mL2And (4) nanosheet mother liquor.
In addition, the inventors characterized by transmission electron microscopy, indicating MnO2The synthesis of the nano-sheet is successful, and particularly, refer to the attached figure 3 in the specification.
It is still another object of the present invention to provide the above-mentioned fluorescent probe PBI-DM @ MnO2Specific application of NS in detecting AChE activity in PBS buffer.
It should be noted that the PBI-DM disclosed herein has two positive charges, increasing its water solubility, and has a strong fluorescence signal in a dispersed state, whereas MnO used in the present invention2The nanosheets are negatively charged and can be electrostatically adsorbed with the PBI-DM to induce the aggregation of the PBI-DM, so that the fluorescence of the PBI-DM is quenched. And AChE is capable of hydrolyzing ATCh to produce thiocholine (TCh), which has reducing properties and reduces MnO2Nanosheet of MnO2The nanosheets are decomposed, so that the structure of the assembly is destroyed, and the ordered stacking mode between the PBI-DM is broken and converted into the existence form of the monomer, so that the fluorescence of the solution is recovered.
In particular, the fluorescent probe PBI-DM @ MnO2The specific application of NS in PBS buffer (pH 7.4) for selectively identifying and quantitatively detecting acetylcholinesterase AChE is as follows:
the fluorescent probe PBI-DM @ MnO disclosed in the invention2Adding acetylcholinesterase AChE and NS into PBS buffer solution (pH 7.4)In the corresponding fluorescence spectrum of the aqueous solution after 30min of acetylcholine ATCH reaction, the addition of acetylcholinesterase AChE can obviously enhance the emission peaks at the wavelength of 550nm and 590nm, while the addition of other proteins can obviously enhance the emission peaks of a fluorescence probe PBI-DM @ MnO2The fluorescence of aqueous solutions of NS has little effect, see in particular figure 4 of the description.
In some application scenes, the fluorescent probe PBI-DM @ MnO is further included2The application of NS in detection and screening of acetylcholinesterase AChE inhibitors by taking acetylcholinesterase AChE as a marker.
According to the technical scheme, compared with the prior art, the invention discloses a fluorescent probe for detecting acetylcholinesterase, a synthetic method and an application thereof, and the fluorescent probe has the following excellent characteristics:
(1) the invention respectively synthesizes PBI-DM with two positive charges and negative MnO with higher water solubility through simple and effective reaction2The nano-sheets induce PBI-DM aggregation fluorescence quenching by utilizing electrostatic interaction between the nano-sheets, when AChE is contained in the system, ATCH can be hydrolyzed to generate TCh, and the TCh can reduce MnO2Decomposing the nano-sheets to release the PBI-DM, and finally detecting the AChE activity by using the fluorescence spectrum characteristics of the PBI-DM in the aggregation and dispersion processes;
(2) the synthesized fluorescent probe disclosed by the invention has high sensitivity in a low background detection mode, has a detection limit as low as 0.24mU/mL, and is suitable for detecting AChE activity in a pure water solvent;
(3) the invention discloses that Glutathione (GSH), cysteine (Cys) and homocysteine (Hcy) have little influence on the fluorescence intensity of the detection mode under the condition of low concentration and in the presence of some other proteins, and have high specificity on the detection of acetylcholinesterase AChE;
(4) the sensing detection analysis disclosed by the invention is successfully applied to the screening of the AChE inhibitor, and further proves the sensitivity and accuracy of the synthesized fluorescent probe disclosed by the invention on the detection of the activity of the acetylcholinesterase AChE.
In conclusion, the invention discloses a synthesized fluorescent probe PBI-DM @ MnO2NS can effectively and selectively detect AChE activity and has high sensitivity to AChE; the invention provides a new AChE detection approach, has the advantages of simple preparation, high specificity and the like, and has market application and popularization values in the aspects of clinical diagnosis, biological field and the like.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the provided drawings without creative efforts.
FIG. 1 is a diagram showing the hydrogen nuclear magnetic resonance spectrum of PBI-DM in heavy water in the fluorescent probe of the invention.
FIG. 2 is a nuclear magnetic resonance carbon spectrum of PBI-DM in heavy water in the fluorescent probe of the invention.
FIG. 3 is a drawing showing MnO in the present invention2Nanosheet and fluorescent probe PBI-DM @ MnO2Transmission electron microscopy image of NS.
FIG. 4 is a graph showing fluorescence spectra of the fluorescent probe of the present invention and other protein analytes.
FIG. 5 is a drawing showing MnO in the present invention2Nanosheet and fluorescent probe PBI-DM @ MnO2NS Zeta potential characterization chart.
FIG. 6 is a graph showing fluorescence spectra of the fluorescent probe of the present invention under different concentrations of AChE.
FIG. 7 is a graph showing the fluorescence spectrum of the interaction of the fluorescent probe of the present invention with GSH, Cys and Hcy.
FIG. 8 is a graph showing fluorescence spectra of the fluorescent probe of the present invention in the presence of AChE inhibitor (tacrine) at various concentrations.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The embodiment of the invention discloses a method based on MnO2The electrostatic interaction between the nanosheets and the PBI-DM, and the novel method for detecting the AChE activity by the fluorescence spectrum characteristics of the PBI in the aggregation and dispersion process provide the probe PBI-DM @ MnO2The synthetic method and the application of NS not only realize the quantitative detection of AChE in the water phase and avoid the interference of other mercapto compounds in the detection process, but also realize the fluorescent probe PBI-DM @ MnO2The NS can be successfully applied to screening of the AChE inhibitor, and has good stability and wide application prospect.
The present invention will be further specifically illustrated by the following examples for better understanding, but the present invention is not to be construed as being limited thereto, and certain insubstantial modifications and adaptations of the invention by those skilled in the art based on the foregoing disclosure are intended to be included within the scope of the invention.
The invention discloses a novel method for detecting the AChE activity of acetylcholinesterase, which is mainly based on MnO2The AChE activity is detected by the electrostatic interaction between the nanosheets and the PBI-DM and the fluorescence spectrum characteristics of the PBI-DM in the aggregation and dispersion processes.
Specifically, the structural formula of PBI-DM in the fluorescent probe is as follows:
the invention also discloses a synthetic method of the fluorescent probe for detecting AChE, which comprises the following steps:
PBS buffer (pH 7.4) was used as a reaction medium, and a DMSO solution of PBI-DM and MnO were added2The nanosheet mother liquor is subjected to electrostatic adsorption and uniformly mixed to obtain the fluorescent probe PBI-DM @ MnO synthesized by the method disclosed by the invention2NS。
In order to further optimize the technical schemeThe concentration of the PBI is 6 x 10-6mol·L-1，MnO2The concentration of the nanosheet mother liquor is 7.65 mug/mL.
To further verify the excellent effects of the present invention, the inventors also conducted the following experiments:
experiment 1: synthesis and structural characterization of PBI
1. Synthesis of PBI-DM
Adding 0.2g of perylene tetracarboxylic anhydride and 0.557mL of N, N-dimethylethylamine into a DMF solvent, adding 0.4g of zinc acetate as a catalyst, heating at 120 ℃, refluxing for reaction overnight, adding secondary water for washing, and drying to obtain a first-step reaction product; adding a toluene solvent into the obtained reaction product, adding methyl iodide, heating and refluxing for 3h, adding diethyl ether to filter and wash the precipitate, and performing suction filtration and vacuum drying to obtain a brownish red product and obtain a target pure product PBI-DM.
2. Test analysis
FIG. 1 shows the luminescent substances PBI-DM1The specific peak value of the HNMR map is as follows:1H NMR(500MHz,DMSO-d6)δ(ppm):8.73(d,J＝8.1Hz,4H),8.46(d,J＝7.9Hz,4H),4.57–4.45(m,4H),3.71(s,4H),3.29(s,18H).
FIG. 2 shows the luminescent substances PBI-DM13C NMR spectrum, specific peak value:13C NMR(126MHz,DMSO-d6)δ(ppm):163.18,134.40,131.40,125.72,124.71,122.83,62.40,53.08,34.29.
the H spectrogram and the C spectrogram of the fluorescent substance PBI-DM prepared by the synthesis method disclosed by the invention are known, and the spectral peaks in the attached drawing correspond to the PBI-DM one by one, so that the synthesis success of the luminous substance PBI-DM can be proved.
Experiment 2: MnO2Synthesis and structural characterization of nanosheets
1、MnO2Synthesis of nanoplatelets
First, a catalyst having a concentration of 0.6 mol.L was prepared-1Tetramethylammonium hydroxide and 3% H2O220mL of mixed aqueous solution; then, the obtained reaction solution was added dropwise to the reaction solution 15s in an amount of 10mL to a concentration of 0.3 mol. L-1MnCl of2In the solution, the solution turns dark brown and is stirred vigorously at room temperatureAt night; finally, centrifuging the obtained reaction product crude product at 2000rpm for 10min, washing the crude product with a large amount of secondary water and methanol, and drying the crude product at 60 ℃ to obtain a target pure product MnO2Nanosheets.
Taking 0.01g of MnO2Dispersing the nano sheet into 40mL of secondary water, performing ultrasonic treatment for 10h, centrifuging at 2000rpm, and leaving supernatant to obtain MnO with the concentration of 2.78mg/mL2And (4) nanosheet mother liquor.
2. Test analysis
MnO can be obviously seen from a transmission electron microscope picture2Sheet structure of (B) indicating MnO2The synthesis of the nano-sheet is successful, and the nano-sheet has good dispersibility, which is shown in figure 3. In addition, MnO can be seen from Zeta potential2The nano-sheet is electronegative, and the Zeta potential reaches-35.4 mV, so that the nano-sheet can be stably dispersed in an aqueous solution, see attached figure 5.
Experiment 3: fluorescent probe PBI-DM @ MnO2Synthesis and structural characterization of NS
1. Fluorescent probe PBI-DM @ MnO2Synthesis of NS
To a 5mL colorimetric tube, 2mL of PBS buffer (pH 7.4) was added as a solvent, and 2.4 μ L of a PBI-DM solution in DMSO (5 × 10) was added thereto-3mol·L-1) Then, 5.5. mu.L of MnO was added2Uniformly mixing the nano-sheet mother solution (2.78mg/mL) to obtain the fluorescent probe PBI-DM @ MnO2NS。
2. Test analysis
MnO can be obviously seen from a transmission electron microscope picture2The PBI-DM loaded by the nanosheets still has a sheet structure and good dispersibility, and is shown in figure 3. In addition, MnO can be seen from Zeta potential2The nanometer sheet is electronegative, after PBI-DM with positive charge is loaded through electrostatic adsorption, the Zeta potential reaches 18.0mV and becomes positive value, which indicates that PBI-DM is successfully adsorbed to MnO2Nano-sheets and can be stably dispersed in an aqueous solution, see figure 5.
Experiment 4: fluorescent probe PBI-DM @ MnO2Specific application of NS in detecting acetylcholinesterase AChE in solvent system
1. Test experiments
Firstly, firstlyAdding 2mL of prepared fluorescent probe PBI-DM @ MnO into the colorimetric tube2NS solution, adding aqueous solution (1 × 10) of Adenosine Triphosphate (ATP), Thrombin (TB), Telomerase (Telomerase), Lysozyme (LZM), thermo-sensitive alkaline phosphatase (TAP), Human Serum Albumin (HSA), Protein Kinase (PKA), inorganic pyrophosphatase (IPPase), glucose (glucose) into each colorimetric tube in turn-2mol·L-1)40 μ L, at a final analyte concentration of 2X 10-4mol·L-1The fluorescence spectrum was measured after mixing, see FIG. 4.
2. Test analysis
In the fluorescence spectrum, the addition of acetylcholinesterase AChE leads PBI-DM @ MnO2The NS solution has a significantly enhanced emission peak at 550nm, while the addition of other proteins to PBI-DM @ MnO2The fluorescence of the aqueous NS solution had no significant effect. Therefore, the fluorescent probe PBI-DM @ MnO prepared by the invention2NS can realize the selective recognition of AChE in a pure water system.
Experiment 5: fluorescent probe PBI-DM @ MnO2Determination of minimum detection limit of NS on AChE quantitative detection
First, 200. mu.L of PBS buffer (pH 7.4) was added with acetylcholinesterase AChE at different concentrations and 3.2. mu.L of an aqueous acetylcholine ATCH solution (1X 10)-2mol·L-1) Reacting at 37 ℃ for 30min, and adding 1.8mL of fluorescent probe PBI-DM @ MnO2The NS solution was mixed well and the fluorescence spectrum was measured, see FIG. 6.
The fluorescence emission spectrum is utilized, the lowest detection limit of the fluorescent probe to AChE is obtained by 3.29 sigma/k calculation, and the result shows that the fluorescent probe PBI-DM @ MnO is2The detection sensitivity of NS on AChE is very high, which indicates that the probe has potential application value in the aspect of efficiently detecting AChE in pure water solution.
Experiment 6: fluorescent probe PBI-DM @ MnO2Anti-interference ability of NS to other sulfhydryl compounds
1. Test experiments
In PBI-DM @ MnO2Different concentrations of CSH, Cys, Hcy, and TCh, a product of AChE and ATCh, were added to NS in PBS buffer (pH 7.4) and measured separatelyFluorescence spectra, see in particular FIG. 7.
2. Test analysis
Taking the fluorescence intensity at 550nm as a reference, the common three sulfhydryl compounds of CSH, Cys and Hcy in the human body under the condition of low concentration to PBI-DM @ MnO2The fluorescence intensity of NS has almost no influence and has high sensitivity on the detection of TCh, which shows that the probe can avoid the interference of CSH, Cys and Hcy on the determination in a certain range, and the activity of AChE can be effectively determined.
Experiment 7: fluorescent probe PBI-DM @ MnO2Specific application of NS in AChE inhibitor screening
First, to 200 μ L of PBS buffer (pH 7.4), 5.0 μ L of AChE aqueous solution (10U/mL) and 3.2 μ LATCh aqueous solution (1 × 10) were added-2mol·L-1) And tacrine aqueous solutions of different concentrations (final concentration 10 each)-11、10-10、10-9、10-8、10-7、10-6、10-5、10-4mol·L-1) Reacting at 37 ℃ for 30min, and adding 1.8mL of PBI-DM @ MnO2And (3) evenly mixing the NS solution and measuring the fluorescence spectrum of the NS solution. Calculating to obtain IC50195.1nM, consistent with the literature. The above results show that the probe PBI-DM @ MnO2NS may be suitable for AChE inhibitor screening, see figure 8.
In summary, the PBI-DM disclosed by the invention has two positive charges, the water solubility of the PBI-DM is increased, and the PBI-DM has strong fluorescence signals in a dispersed state, while MnO used in the invention is MnO2The nanoplatelets are negatively charged and can be electrostatically adsorbed with the PBI-DM to induce the aggregation of the PBI-DM and quench the initial fluorescence, so that the PBI-DM has a low optical background. And AChE can hydrolyze ATCH to produce TCh, which has reducing property and can reduce MnO2Nanosheet of MnO2The nanosheets are decomposed, so that the structure of the assembly is destroyed, and the ordered stacking mode between the PBI-DM is broken and converted into the existence form of the monomer, so that the fluorescence of the solution is recovered.
And the fluorescent probe PBI-DM @ MnO2The detection of AChE by NS has a low detection limit of 0.24mU/mL, and can avoid low concentration of other sulfhydryl compoundsThe interference of (2) has higher selectivity; and the characterization through a transmission electron microscope, a Zeta potential and the like proves that the fluorescent probe PBI-DM @ MnO is2The NS is successfully synthesized and has excellent dispersibility in aqueous solution; meanwhile, the invention also verifies that the fluorescent probe PBI-DM @ MnO2The feasibility of NS for AChE inhibitor screening means that the invention has great potential in the field of clinical diagnosis.
The previous description of the disclosed embodiments and examples is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (10)
1. The cationic derivative of perylene tetracarboxylic diimide is characterized in that the structural formula of the cationic derivative PBI-DM is as follows:
2. the method for synthesizing cationic derivatives of perylene tetracarboxylic diimides as claimed in claim 1, which comprises the following steps:
(1) dimethyl formamide is used as a reaction medium, perylene tetracarboxylic anhydride and N, N-dimethylethylamine in a molar ratio of 1:10 are used as substrates, zinc acetate is used as a catalyst, and after heating reflux reaction is carried out overnight, washing and drying are carried out to obtain a reaction product I;
(2) adding a toluene solvent into the reaction product I obtained in the step (1), taking methyl iodide as a reaction substrate, and heating and refluxing for reaction for 3 hours to obtain a reaction product II;
(3) and (3) adding diethyl ether into the reaction product II obtained in the step (2) to filter and wash the precipitate, and performing suction filtration and vacuum drying to obtain a brownish red product, namely the target pure product of the cationic derivative PBI-DM of the perylene tetracarboxylic diimide.
3. MnO (MnO)2The synthesis method of the nanosheet mother liquor is characterized by comprising the following steps:
(A) mixing tetramethylammonium hydroxide and H2O2The mixed solution is dripped into MnCl2Stirring the solution vigorously overnight to obtain a reaction product;
(B) centrifuging, washing and drying the reaction product obtained in the step (A) to obtain a pure product MnO2Nanosheets;
(C) MnO is carried out on the pure product obtained in the step (B)2Dispersing the nano sheet into water, performing ultrasonic treatment for 10h, centrifuging, and leaving supernatant to obtain MnO2And (4) nanosheet mother liquor.
4. The MnO of claim 32The synthesis method of the nanosheet mother liquor is characterized in that in the step (A), tetramethylammonium hydroxide and H2O2Is 6: 10, tetramethylammonium hydroxide and the MnCl in the mixed solution2The molar ratio of the solution is 2: 1; and the dropping speed of the mixed solution in the step (A) is 1.5-2 mL/s, and the reaction temperature is 20-30 ℃.
5. The MnO of claim 32The synthesis method of the nanosheet mother liquor is characterized in that the centrifugation rates in the step (B) and the step (C) are both 1500-2000 rpm, the centrifugation time is 10-15 min, and the drying temperature is 60-80 ℃.
6. A fluorescent probe for detecting the activity of acetylcholinesterase is characterized in that the fluorescent probe is MnO loaded with PBI-DM2Nanosheet with molecular formula of PBI-DM @ MnO2NS; and the fluorescent probe is based on MnO as defined in claim 32Electrostatic interaction between nanoplatelets and the PBI-DM of claim 1, and the sameAnd (3) the fluorescence spectrum characteristics of the PBI-DM in the aggregation and dispersion process so as to realize the detection of the acetylcholinesterase activity.
7. A method for synthesizing a fluorescent probe for detecting acetylcholinesterase activity according to claim 6, wherein said method comprises the following steps:
(a) adding PBS buffer solution into a colorimetric tube as a solvent;
(b) adding a dimethyl sulfoxide solution of the cationic derivative of perylene tetracarboxylic diimide and the MnO to the solvent of step (a)2Obtaining a fluorescent probe PBI-DM @ MnO used for detecting the activity of acetylcholinesterase by electrostatic adsorption of nanosheet mother liquor2NS。
8. The method as claimed in claim 7, wherein the concentration of the perylene tetracarboxylic diimide cation derivative in the step (b) is 6 x 10-6mol·L-1Said MnO being2The concentration of the nanosheet mother liquor is 7.65 mug/mL.
9. Use of the fluorescent probe for detecting acetylcholinesterase activity according to claim 6 or the fluorescent probe synthesized by the method according to claim 7 in a solvent system to selectively recognize acetylcholinesterase.
10. The use of the fluorescent probe for detecting acetylcholinesterase activity according to claim 9, further comprising the use of the fluorescent probe in the detection and screening of acetylcholinesterase inhibitors using acetylcholinesterase as a marker.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20150243869A1 (en) * | 2014-02-20 | 2015-08-27 | The Regents Of The University Of California | Self doping materials and methods |
| CN105130988A (en) * | 2015-07-31 | 2015-12-09 | 南京邮电大学 | Ion type perylene imide material, preparation method, and organic storage device thereof |
| CN105548167A (en) * | 2015-12-29 | 2016-05-04 | 安徽师范大学 | Manganese dioxide sheet mimic enzyme sensor and preparation method thereof as well as T4PNK detection method |
| CN110156781A (en) * | 2019-06-21 | 2019-08-23 | 河南大学 | Application of the tetracarboxylic acid dianhydride amidation compound in anti-Staphylococcus aureus |
-
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20150243869A1 (en) * | 2014-02-20 | 2015-08-27 | The Regents Of The University Of California | Self doping materials and methods |
| CN105130988A (en) * | 2015-07-31 | 2015-12-09 | 南京邮电大学 | Ion type perylene imide material, preparation method, and organic storage device thereof |
| CN105548167A (en) * | 2015-12-29 | 2016-05-04 | 安徽师范大学 | Manganese dioxide sheet mimic enzyme sensor and preparation method thereof as well as T4PNK detection method |
| CN110156781A (en) * | 2019-06-21 | 2019-08-23 | 河南大学 | Application of the tetracarboxylic acid dianhydride amidation compound in anti-Staphylococcus aureus |
Non-Patent Citations (1)
| Title |
|---|
| T. DELIGEORGIEV等: "《Synthesis and Properties of Fluorescent》", 《DYES AND PIGMENTS》 * |
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