CN118221582A - Ratio type fluorescent probe of butyrylcholine esterase and synthetic method and application thereof - Google Patents
Ratio type fluorescent probe of butyrylcholine esterase and synthetic method and application thereof Download PDFInfo
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- 102100032404 Cholinesterase Human genes 0.000 title claims abstract description 57
- 239000007850 fluorescent dye Substances 0.000 title claims abstract description 52
- 101710083761 Cholinesterase Proteins 0.000 title claims abstract description 19
- 238000010189 synthetic method Methods 0.000 title description 2
- 108010053652 Butyrylcholinesterase Proteins 0.000 claims abstract description 38
- 239000000523 sample Substances 0.000 claims abstract description 28
- 239000000575 pesticide Substances 0.000 claims abstract description 26
- 230000000694 effects Effects 0.000 claims abstract description 20
- KXDHJXZQYSOELW-UHFFFAOYSA-M Carbamate Chemical compound NC([O-])=O KXDHJXZQYSOELW-UHFFFAOYSA-M 0.000 claims abstract description 11
- 238000004445 quantitative analysis Methods 0.000 claims abstract description 8
- 238000000034 method Methods 0.000 claims description 15
- 238000006243 chemical reaction Methods 0.000 claims description 9
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 5
- 150000001875 compounds Chemical class 0.000 claims description 5
- ZOOSILUVXHVRJE-UHFFFAOYSA-N cyclopropanecarbonyl chloride Chemical compound ClC(=O)C1CC1 ZOOSILUVXHVRJE-UHFFFAOYSA-N 0.000 claims description 4
- 238000011534 incubation Methods 0.000 claims description 4
- 239000003960 organic solvent Substances 0.000 claims description 4
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- 229910000024 caesium carbonate Inorganic materials 0.000 claims description 2
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims 3
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- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 claims 2
- VZGDMQKNWNREIO-UHFFFAOYSA-N tetrachloromethane Chemical compound ClC(Cl)(Cl)Cl VZGDMQKNWNREIO-UHFFFAOYSA-N 0.000 claims 2
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- UIIMBOGNXHQVGW-DEQYMQKBSA-M Sodium bicarbonate-14C Chemical compound [Na+].O[14C]([O-])=O UIIMBOGNXHQVGW-DEQYMQKBSA-M 0.000 claims 1
- 125000001559 cyclopropyl group Chemical group [H]C1([H])C([H])([H])C1([H])* 0.000 claims 1
- 230000005284 excitation Effects 0.000 claims 1
- PSHKMPUSSFXUIA-UHFFFAOYSA-N n,n-dimethylpyridin-2-amine Chemical compound CN(C)C1=CC=CC=N1 PSHKMPUSSFXUIA-UHFFFAOYSA-N 0.000 claims 1
- 229910000027 potassium carbonate Inorganic materials 0.000 claims 1
- 238000002360 preparation method Methods 0.000 claims 1
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 claims 1
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- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims 1
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- QGLZXHRNAYXIBU-WEVVVXLNSA-N aldicarb Chemical compound CNC(=O)O\N=C\C(C)(C)SC QGLZXHRNAYXIBU-WEVVVXLNSA-N 0.000 description 7
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
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- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical compound [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
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- WYMSBXTXOHUIGT-UHFFFAOYSA-N paraoxon Chemical compound CCOP(=O)(OCC)OC1=CC=C([N+]([O-])=O)C=C1 WYMSBXTXOHUIGT-UHFFFAOYSA-N 0.000 description 3
- 238000012271 agricultural production Methods 0.000 description 2
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- DUEPRVBVGDRKAG-UHFFFAOYSA-N carbofuran Chemical compound CNC(=O)OC1=CC=CC2=C1OC(C)(C)C2 DUEPRVBVGDRKAG-UHFFFAOYSA-N 0.000 description 2
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- LCCNCVORNKJIRZ-UHFFFAOYSA-N parathion Chemical compound CCOP(=S)(OCC)OC1=CC=C([N+]([O-])=O)C=C1 LCCNCVORNKJIRZ-UHFFFAOYSA-N 0.000 description 2
- 229910001392 phosphorus oxide Inorganic materials 0.000 description 2
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- VSAISIQCTGDGPU-UHFFFAOYSA-N tetraphosphorus hexaoxide Chemical compound O1P(O2)OP3OP1OP2O3 VSAISIQCTGDGPU-UHFFFAOYSA-N 0.000 description 2
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 1
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- 239000005944 Chlorpyrifos Substances 0.000 description 1
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- 206010059866 Drug resistance Diseases 0.000 description 1
- 239000005949 Malathion Substances 0.000 description 1
- LSDPWZHWYPCBBB-UHFFFAOYSA-N Methanethiol Chemical compound SC LSDPWZHWYPCBBB-UHFFFAOYSA-N 0.000 description 1
- 239000005916 Methomyl Substances 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
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- CVXBEEMKQHEXEN-UHFFFAOYSA-N carbaryl Chemical compound C1=CC=C2C(OC(=O)NC)=CC=CC2=C1 CVXBEEMKQHEXEN-UHFFFAOYSA-N 0.000 description 1
- 229960005286 carbaryl Drugs 0.000 description 1
- 231100000504 carcinogenesis Toxicity 0.000 description 1
- SBPBAQFWLVIOKP-UHFFFAOYSA-N chlorpyrifos Chemical compound CCOP(=S)(OCC)OC1=NC(Cl)=C(Cl)C=C1Cl SBPBAQFWLVIOKP-UHFFFAOYSA-N 0.000 description 1
- 229940048961 cholinesterase Drugs 0.000 description 1
- 231100000739 chronic poisoning Toxicity 0.000 description 1
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- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- OEBRKCOSUFCWJD-UHFFFAOYSA-N dichlorvos Chemical compound COP(=O)(OC)OC=C(Cl)Cl OEBRKCOSUFCWJD-UHFFFAOYSA-N 0.000 description 1
- 229950001327 dichlorvos Drugs 0.000 description 1
- JXSJBGJIGXNWCI-UHFFFAOYSA-N diethyl 2-[(dimethoxyphosphorothioyl)thio]succinate Chemical compound CCOC(=O)CC(SP(=S)(OC)OC)C(=O)OCC JXSJBGJIGXNWCI-UHFFFAOYSA-N 0.000 description 1
- MCWXGJITAZMZEV-UHFFFAOYSA-N dimethoate Chemical compound CNC(=O)CSP(=S)(OC)OC MCWXGJITAZMZEV-UHFFFAOYSA-N 0.000 description 1
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- NNKVPIKMPCQWCG-UHFFFAOYSA-N methamidophos Chemical compound COP(N)(=O)SC NNKVPIKMPCQWCG-UHFFFAOYSA-N 0.000 description 1
- UHXUZOCRWCRNSJ-QPJJXVBHSA-N methomyl Chemical compound CNC(=O)O\N=C(/C)SC UHXUZOCRWCRNSJ-QPJJXVBHSA-N 0.000 description 1
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- BULVZWIRKLYCBC-UHFFFAOYSA-N phorate Chemical compound CCOP(=S)(OCC)SCSCC BULVZWIRKLYCBC-UHFFFAOYSA-N 0.000 description 1
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- NFACJZMKEDPNKN-UHFFFAOYSA-N trichlorfon Chemical compound COP(=O)(OC)C(O)C(Cl)(Cl)Cl NFACJZMKEDPNKN-UHFFFAOYSA-N 0.000 description 1
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- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention provides a ratio type fluorescent probe of butyrylcholine esterase, a synthesis method and application thereof, wherein the fluorescent probe utilizes the fluorescence intensity ratio of double emission peaks to analyze butyrylcholine esterase activity, and the interference of factors such as probe concentration, microenvironment, light source intensity and the like can be effectively reduced in a self-correcting mode, so that the accuracy of a detection result is high. In addition, the fluorescent probe has high sensitivity to butyrylcholinesterase and is not interfered by coexisting matters such as ions, amino acids, polysaccharides, proteins and the like. The fluorescent probe has simple synthesis steps, is easy to separate and purify, has good stability, is suitable for batch production, and is favorable for commercialized popularization and application; the fluorescent probe can be applied to the detection of butyrylcholine esterase activity, can also be applied to the quantitative analysis of organophosphorus pesticides and carbamate pesticides, and has great market application and popularization value in the pesticide residue detection field.
Description
Technical Field
The invention relates to the technical field of pesticide detection, in particular to a ratio type fluorescent probe of butyrylcholine esterase and a synthesis method and application thereof.
Background
The problem of pesticide residues in food has been a focus of attention. With the progress of scientific technology, the agricultural production scale is continuously enlarged, the pesticide usage amount is continuously increased, and the problem of pesticide residue is increasingly serious. Eating a large amount of food with excessive pesticide residues easily causes acute poisoning symptoms such as emesis, abdominal pain or nerve dysfunction, and the serious patients can cause death; chronic poisoning is easy to occur even if the food with low pesticide residue is eaten for a long time; in addition, the phenomenon of teratogenesis and carcinogenesis of pesticides is also common. At present, pesticides used in agricultural production are various, and organophosphorus and carbamate pesticides are the two most commonly used pesticides, so rapid and accurate detection of the two pesticides has become a focus of attention in the field of food safety.
Currently, the main methods for detecting pesticide residues in agricultural products are chromatography, immunological methods, biological detection methods and enzyme inhibition methods. Although methods for detecting pesticide residues are various, various drawbacks exist. The chromatographic method has high sensitivity, but the instrument operation and the sample pretreatment are complex and time-consuming, and cannot be used for real-time rapid detection of actual samples; the immunological method has mature technology and wide application, but the interference of various pesticides can influence the detection result; the biological detection method does not need to carry out pretreatment on the sample, but has the defects of rough results, easy generation of drug resistance of organisms used and the like. The enzyme inhibition method based on the inhibition of cholinesterase by organophosphorus or carbamate pesticides has the advantages of simple operation, rapid reaction and the like, and has become a common method for rapidly detecting pesticide residues. However, most reported fluorescent probes based on the enzyme inhibition method have the defects of serious interference by mercaptan, low sensitivity, complex operation and the like.
Therefore, how to establish a detection method which has strong anti-interference capability and simple and convenient operation and can monitor organophosphorus pesticides and carbamate pesticides with high sensitivity based on an enzyme inhibition method is a problem which needs to be solved by the technicians in the field.
Disclosure of Invention
The invention aims to provide a ratio type fluorescent probe of butyrylcholine esterase as well as a synthesis method and application thereof, so as to solve the problems in the background technology.
According to an object of the present invention, there is provided the above ratio-type fluorescent probe of butyrylcholinesterase having a structure represented by formula (1)
According to another object of the present invention, there is provided a method for synthesizing the above ratio-type fluorescent probe of butyrylcholinesterase, wherein a compound represented by formula (2) is reacted with cyclopropylcarbonyl chloride in an organic solvent in the presence of a base;
according to a third object of the present invention, there is provided the use of the above ratio-type fluorescent probe for detecting butyrylcholinesterase activity.
Further, the invention provides a detection method of butyrylcholinesterase activity, wherein the method comprises the following steps: the fluorescent probe is added into a sample to be detected, and incubation is carried out at a certain temperature, so that the fluorescent probe reacts with butyrylcholine esterase in the sample to be detected and then the cyclopropane formyl group is removed, and a compound shown in a formula (2) is generated; the ratio of the fluorescence intensities of the two emission peaks of the fluorescent probe is in proportional relation with the activity concentration of butyrylcholinesterase, so that the activity of butyrylcholinesterase can be quantified.
Further, the invention provides application of the ratio type fluorescent probe in monitoring inhibition of organophosphorus pesticides and carbamate pesticides on butyrylcholine esterase. The organophosphorus pesticide comprises parathion, phorate, trichlorfon, methamidophos, dichlorvos, malathion, chlorpyrifos, dimethoate and parathion; the carbamate pesticide comprises: carbaryl, aldicarb, carbofuran, methomyl.
The beneficial effects of the invention are as follows:
the fluorescence probe analyzes butyrylcholine esterase activity by utilizing the fluorescence intensity ratio of the double emission peaks, and can effectively reduce the interference of factors such as probe concentration, microenvironment, light source intensity and the like in a self-correcting mode, so that the accuracy of a detection result is high. In addition, the fluorescent probe has high sensitivity to butyrylcholinesterase and is not interfered by coexisting matters such as ions, amino acids, polysaccharides, proteins and the like. The fluorescent probe has the advantages of simple synthesis steps, easy separation and purification, good stability, suitability for mass production and contribution to commercialization popularization and application.
The fluorescent probe can be applied to detection of butyrylcholine esterase activity, can be applied to quantitative analysis of organophosphorus pesticides and carbamate pesticides, and has great market application and popularization value in rapid detection of pesticide residues.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the description of the embodiments or the prior art will be briefly described, and it is obvious that the drawings in the description below are some embodiments of the present invention, and other drawings can be obtained according to the drawings without inventive effort for a person skilled in the art.
FIG. 1 is a 1 H NMR spectrum of a fluorescent probe of the invention.
FIG. 2 is a 13 C NMR spectrum of a fluorescent probe of the invention.
FIG. 3 shows the absorption spectrum (a) and the fluorescence spectrum (b) of the fluorescent probe of the present invention after reaction with butyrylcholinesterase.
FIG. 4 is a linear relationship (b) between the fluorescence spectrum of the fluorescent probe of the present invention and the ratio of fluorescence intensity of the butyrylcholinesterase enzyme response (a) and its dual emission peak and the activity concentration of butyrylcholinesterase enzyme.
FIG. 5 shows fluorescence spectra (a) and inhibition efficiency curves (b) of the fluorescent probe and butyrylcholine esterase according to the invention after addition of p-oxyphosphorus reaction.
FIG. 6 shows fluorescence spectra (a) and inhibition efficiency curves (b) of the fluorescent probe of the present invention reacted with butyrylcholinesterase after addition of aldicarb.
Detailed Description
The embodiment of the invention discloses a high-selectivity, high-sensitivity and ratio type fluorescent probe for detecting butyrylcholine esterase activity, and a synthesis method and application thereof.
The present invention will be further specifically illustrated by the following examples, which are not to be construed as limiting the invention, but rather as falling within the scope of the present invention, for some non-essential modifications and adaptations of the invention that are apparent to those skilled in the art based on the foregoing disclosure.
The invention discloses a ratio type fluorescent probe for measuring butyrylcholine esterase activity, which has the structural formula:
the technical scheme of the invention will be further described below with reference to specific embodiments.
Example 1: synthesis of fluorescent probes
The synthesis steps are as follows: compound 1 (0.14 g,0.5 mmol) was dissolved in 20mL of CH 2Cl2, cesium carbonate (1.10 g,3.0 mmol) was added, and after stirring in an ice-water bath for 30 minutes, cyclopropylcarbonyl chloride (276. Mu.L, 3.0 mmoL) was added dropwise and stirring was continued for 8 hours, after which the reaction was stopped. The reaction solution is decompressed and concentrated to remove the solvent, and the residue is separated by a column (silica gel, 200-300 meshes, eluent is methylene dichloride/methanol mixed solution with the volume ratio of 20:1) to obtain the ratio type fluorescent probe for detecting butyrylcholine esterase activity. The synthetic route of the fluorescent probe is as follows:
FIG. 1 is 1H NMR(500MHz,DMSO-d6) spectrum of a ratio-type fluorescent probe ：δppm 8.66(dd,J＝7.3,1.1Hz,1H),8.63(d,J＝8.0Hz,1H),8.41(dd,J＝8.4,1.2Hz,1H),7.92(dd,J＝8.5,7.3Hz,1H),7.67(d,J＝8.1Hz,1H),4.56(t,J＝5.8Hz,2H),3.56(t,J＝5.8Hz,2H),3.05(s,6H),2.17(tt,J＝7.6,5.4Hz,1H),1.28–1.23(m,4H).
FIG. 2 is 13C NMR(126MHz,DMSO-d6) spectrum of a ratio-type fluorescent probe ：δppm 173.03,164.55,164.00,152.33,131.78,131.60,129.24,128.16,127.29,125.43,122.38,119.71,119.63,56.21,42.89,35.27,12.23,8.83.
1 The H NMR and 13 C NMR spectra correspond to the chemical structure of the ratio-type fluorescent probe, indicating that the probe synthesis was successful.
In order to further verify the excellent effect of the fluorescent probe synthesized by the present disclosure in detecting butyrylcholinesterase (BChE) activity, the following experimental operations were also performed:
experiment 1: test of ability of fluorescent Probe to react with butyrylcholinesterase in buffer solution
(1) The fluorescent probe prepared in example 1 was prepared as a DMSO stock solution (at a concentration of 1 mM), added to an EP tube, and then a PBS buffer (at a concentration of 200mM, pH 7) and a stock solution of BChE (at a concentration of 10U/mL) were sequentially added to the tube. The total volume of the solution was 1mL, the final concentration of the probe was 10. Mu.M, the final concentration of PBS was 20mM, and the final concentration of BChE was 0.2U/mL. The above solution was left in a constant temperature incubator at 37℃for 1 hour, and then its absorption spectrum and fluorescence spectrum were measured.
FIG. 3 shows the absorption spectrum (a) and fluorescence emission spectrum (b) before and after the reaction of the probe with BChE. After the probe reacted with BChE, its absorbance peak red shifted from 334nm to 446nm (panel a in fig. 3); the fluorescence intensity at 445nm was significantly reduced and the fluorescence intensity at 560nm was significantly increased to form a new emission peak (panel b in fig. 3). The results indicate that the probe has a typical two-channel fluorescent response to BChE.
(2) Series of EP tubes were taken and probe DMSO stock, deionized water, PBS buffer and stock of BChE of different volumes were added separately. The total volume of the solution was 1mL, the final concentration of the probe was 10. Mu.M, the final concentration of PBS was 20mM, and the final concentration of BChE was 0-0.6U/mL. The above solution was left in a constant temperature incubator at 37℃for 1 hour, and then its fluorescence spectrum was measured.
Fig. 4 is a titration experiment of probe versus BChE concentration. As the BChE concentration was gradually increased to 0.6U/mL, the fluorescence intensity of the probe at the 445nm emission peak was gradually decreased, but the fluorescence intensity at the 560nm emission peak was gradually increased (panel a in FIG. 4). The fluorescence intensity ratio F 560/F445 of the probe at the two emission peaks and BChE concentration show a good linear relationship (R 2 = 0.9958) in the range of 0-0.16u/mL (panel b in fig. 4), indicating that the probe can accurately quantify BChE activity.
Experiment 2: quantitative analysis of organophosphorus pesticides by fluorescent probe
The fluorescence spectrum changes were detected after pretreatment with various concentrations of paraoxone (paraxon, organophosphorus pesticides) with BChE (0.5U/mL) for 20 min, followed by incubation with probe (10 μm) for 1 hour at 37 ℃.
FIG. 5 is a fluorescence spectrum of probe after reaction with organophosphorus pretreated BChE. As the concentration of paraoxon gradually increased to 120ng/mL, the fluorescence intensity of the probe at 560nm gradually decreased (panel a in fig. 5), indicating a significant inhibition of BChE by paraoxon. The inhibition rate and the logarithm of the concentration of phosphorus oxide (40 ng/mL is less than or equal to C paraxon and less than or equal to 90 ng/mL) show good linear relation (R 2 = 0.9922) (b graph in fig. 5), and the method is suitable for quantitative analysis of phosphorus oxide.
Experiment 3: quantitative analysis of carbamate pesticides by fluorescent probe
The fluorescence spectrum changes were detected after pretreatment with various concentrations of aldicarb (aldicarb, carbamate pesticide) with BChE (0.5U/mL) for 20 min, followed by incubation with probe (10 μm) for 1 hour at 37 ℃.
From the fluorescence spectrum of the probe after reaction with carbamate pre-treated BChE, it can be seen that as the concentration of aldicarb gradually increases to 5000ng/mL, the fluorescence intensity of the probe at 560nm gradually decreases (fig. 6, panel a), indicating that aldicarb has a significant inhibitory effect on BChE. The inhibition rate and the logarithm of carbofuran concentration (less than or equal to 1200 ng/mL) show good linear relation (R 2 = 0.9990) (b graph in figure 6), and are suitable for quantitative analysis of aldicarb.
In summary, the invention provides a butyrylcholine esterase ratio type fluorescent probe with high sensitivity, high selectivity and quick response, a synthesis method and application thereof, and establishes a high-efficiency enzyme inhibition rate method for detecting organophosphorus pesticides and carbamate pesticides.
The fluorescence probe provided by the invention utilizes the fluorescence intensity ratio of the double emission peaks to analyze butyrylcholine esterase activity, and can effectively reduce the interference of factors such as probe concentration, microenvironment, light source intensity and the like in a self-correction mode, so that the accuracy of a detection result is high. In addition, the fluorescent probe has high sensitivity to butyrylcholinesterase and is not interfered by coexisting matters such as ions, amino acids, polysaccharides, proteins and the like. The fluorescent probe has the advantages of simple synthesis steps, easy separation and purification, good stability, suitability for mass production and contribution to commercialization popularization and application.
The fluorescent probe can be applied to the detection of butyrylcholine esterase activity, can also be applied to the quantitative analysis of organophosphorus pesticides and carbamate pesticides, and has great market application and popularization value in the pesticide residue detection field.
It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. The present embodiments are, therefore, to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.
Claims (7)
1. A ratio-type fluorescent probe of butyrylcholinesterase, which is characterized by having the structural formula:
2. the method for preparing a ratio-type fluorescent probe of butyrylcholinesterase according to claim 1, comprising the steps of: reacting a compound shown in a formula (2) with cyclopropylcarbonyl chloride in an organic solvent in the presence of a base;
3. The production method according to claim 2, wherein the amount of the cyclopropylcarbonyl chloride is 1 to 6 mmol, the amount of the base is 1 to 12 mmol, and the amount of the organic solvent is 20 to 60 ml, relative to 1 mmol of the structural compound represented by formula (2); the base is at least one of cesium carbonate, potassium carbonate, sodium bicarbonate, pyridine, piperazine, triethylamine and N, N-dimethylaminopyridine; the organic solvent is selected from at least one of dichloromethane, chloroform, carbon tetrachloride, tetrahydrofuran, acetonitrile, acetone, N-dimethylformamide and 1, 2-dichloroethane.
4. A method of preparation according to claim 3, the reaction conditions comprising: the temperature is between 20 ℃ below zero and 60 ℃; the reaction time is as follows: and 1-12 hours.
5. The use of a fluorescent probe according to claim 1, for detecting butyrylcholinesterase activity.
6. The use of a fluorescent probe as claimed in claim 5, comprising the steps of: adding the fluorescent probe into a sample to be detected, and incubating at a certain temperature to enable the fluorescent probe to react with butyrylcholine esterase in the sample to be detected and then removing a cyclopropane group to generate a compound shown in a formula (2); the ratio of the fluorescence intensities of the two emission peaks of the fluorescent probe is in proportional relation with the activity concentration of butyrylcholinesterase, so that the activity of butyrylcholinesterase can be quantified; the incubation conditions included: the temperature is 10-50deg.C, pH is 3-9, and the time is 5-120min; the excitation wavelength of the fluorescent probe is 300-500nm, and the emission wavelength is 400-800nm.
7. The use of a fluorescent probe according to claim 1, characterized in that it is applied to quantitative analysis of organophosphorus pesticides and carbamate pesticides.
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| CN202410213040.8A CN118221582A (en) | 2024-02-27 | 2024-02-27 | Ratio type fluorescent probe of butyrylcholine esterase and synthetic method and application thereof |
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