CN115785952B - Light-sensitive peptide element for fluorescent sensing material, preparation method of light-sensitive peptide element, fluorescent sensing material, preparation method of fluorescent sensing material and application of fluorescent sensing material - Google Patents
Light-sensitive peptide element for fluorescent sensing material, preparation method of light-sensitive peptide element, fluorescent sensing material, preparation method of fluorescent sensing material and application of fluorescent sensing material Download PDFInfo
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Abstract
The invention relates to the technical field of food safety and pesticide detection nanometer, and discloses a light-sensitive peptide element for a fluorescent sensing material, a preparation method thereof, the fluorescent sensing material, and a preparation method and application thereof. The light-sensitive peptide element comprises a peptide-based material and metal particles bridged on the peptide-based material; the peptidyl material is provided by at least one of diphenyl alanine, carbon-terminally amidated diphenyl alanine, di-histidine and phenylalanine-histidine; the metal particles are provided by a metal ion source. The fluorescent sensing material prepared by using the light-sensitive peptide element has the advantages of good selective recognition, high detection sensitivity, quick response and strong adsorption capacity on cartap, and has good practical value.
Description
Technical Field
The invention relates to the technical field of food safety and pesticide detection nanometer, in particular to a light-sensitive peptide element for a fluorescent sensing material, a method for preparing the light-sensitive peptide element for the fluorescent sensing material, the light-sensitive peptide element for the fluorescent sensing material prepared by the method, the fluorescent sensing material, a method for preparing the fluorescent sensing material, the fluorescent sensing material prepared by the method and application of the fluorescent sensing material in detecting cartap.
Background
Nereistoxin is a natural toxin isolated from the worm, isoppodosoma (Lumbriconereis heteroopoda), a marine annelid. As an acetylcholine antagonist, na and K can be inhibited from being conducted through the endplate membrane, thereby causing disturbance of the central nervous system and achieving the insecticidal effect.
Cartap is a bionic pesticide derived from cartap, has high-efficiency insecticidal capability, and is widely applied to modern agriculture for keeping harvest and increasing yield. However, excessive use and improper application can cause residues in vegetable foods, and after conversion into nereistoxin or dihydronereistoxin, the nereistoxin is transported and accumulated in human bodies through food chains, thus causing safety risks for human health. And the total death rate caused by the residual non-productive nereistoxin pesticides in the environment is as high as 11.6% -14.0%. Therefore, the method has practical and important significance for quantitative detection of the nereistoxin pesticide residues in the vegetable foods from the environment.
At present, the quantitative detection of cartap residue is difficult to meet the requirements of standard limit and actual detection due to the higher detection limit of high performance liquid chromatography, so that the quantitative detection is mainly dependent on a large-scale precise instrument of liquid chromatography tandem mass spectrometry. The large-scale precise instrument provides high sensitivity and accuracy, and also has a series of defects which are difficult to ignore, such as expensive instrument cost, higher detection cost, required professionals and incapability of carrying out on-site timely detection.
Disclosure of Invention
The invention aims to provide a light-sensitive peptide element capable of being used for a fluorescent sensing material and a preparation method thereof, and the fluorescent sensing material containing the light-sensitive peptide element can be used for specifically detecting cartap.
The second purpose of the invention is to provide a fluorescent sensing material, a preparation method and application thereof, and the fluorescent sensing material provided by the invention has the advantages of good selective recognition, high detection sensitivity, quick response and strong adsorption capacity on cartap.
In order to achieve the above object, a first aspect of the present invention provides a light-sensitive peptide element for a fluorescent sensing material, the light-sensitive peptide element comprising a peptide-based material and metal particles bridged on the peptide-based material; the peptidyl material is provided by at least one of diphenyl alanine, carbon-terminally amidated diphenyl alanine, di-histidine and phenylalanine-histidine; the metal particles are provided by a metal ion source.
In a second aspect, the present invention provides a method of preparing a light-sensitive peptide element for use in a fluorescent sensing material, the method comprising: self-assembling a metal ion source and a peptide-based material in the presence of a solvent I; the peptide-based material is at least one selected from the group consisting of diphenylalanine, carbon-terminally amidated diphenylalanine, di-histidine and phenylalanine-histidine.
In a third aspect, the present invention provides a light-sensitive peptide element for a fluorescent sensing material prepared by the method described in the second aspect.
In a fourth aspect, the present invention provides a method of preparing a fluorescent sensing material, the method comprising: covalently assembling the rigid component with the light-sensitive peptide element in the presence of solvent II and a catalyst; the light-sensitive peptide element is the light-sensitive peptide element for a fluorescent sensing material according to the first or third aspect; the rigid component is at least one selected from 1, 4-benzene dicarboxaldehyde and 4,4' -biphenyl dicarboxaldehyde.
In a fifth aspect, the present invention provides a fluorescent sensing material prepared by the method according to the fourth aspect.
The sixth aspect of the invention provides an application of the fluorescent sensing material in detecting cartap.
The fluorescent sensing material provided by the invention has the advantages of good selectivity and recognition, quick response, accurate quantification, safety and environmental protection for cartap residue detection.
More specifically, the fluorescent sensing material has the advantages of direct contact sites, cascade response function, rapid diffusion capability and safer and more environment-friendly performance, and realizes rapid response and accurate quantification of cartap residue in a multi-component food matrix.
Additional features and advantages of the invention will be set forth in the detailed description which follows.
Drawings
FIG. 1 is a transmission electron microscope image of the photo-induced peptide element T1 prepared in example 1.
FIG. 2 is a scanning electron microscope image of the fluorescent sensing material L1 prepared in example 1.
Fig. 3 and 4 are fluorescence responses of the light-sensitive peptide element T1 and the fluorescent sensing material L1 in test example 1 to a target at different adsorption times, respectively.
FIG. 5 shows the adsorption capacity of the fluorescent sensor material L1 for a target in test example 2.
Fig. 6 shows the fluorescence intensities of the fluorescent sensing materials L1, L2, L3, and L4.
FIG. 7 shows the adsorption specificity of the fluorescent sensor material L1 to a target.
Detailed Description
The endpoints and any values of the ranges disclosed herein are not limited to the precise range or value, and are understood to encompass values approaching those ranges or values. For numerical ranges, one or more new numerical ranges may be found between the endpoints of each range, between the endpoint of each range and the individual point value, and between the individual point value, in combination with each other, and are to be considered as specifically disclosed herein.
The average particle diameters described in the present invention all represent average diameters.
As previously described, a first aspect of the present invention provides a light-sensitive peptide element for a fluorescent sensing material, the light-sensitive peptide element comprising a peptide-based material and metal particles bridged to the peptide-based material; the peptidyl material is provided by at least one of diphenyl alanine, carbon-terminally amidated diphenyl alanine, di-histidine and phenylalanine-histidine; the metal particles are provided by a metal ion source.
Preferably, the metal ions in the metal ion source are selected from at least one of zinc ions, nickel ions, copper ions and cobalt ions.
More preferably, the metal ion source is at least one selected from zinc chloride, nickel chloride, copper chloride, and cobalt nitrate.
As previously described, a second aspect of the present invention provides a method of preparing a light-sensitive peptide element for use in a fluorescent sensing material, the method comprising: self-assembling a metal ion source and a peptide-based material in the presence of a solvent I; the peptide-based material is at least one selected from the group consisting of diphenylalanine, carbon-terminally amidated diphenylalanine, di-histidine and phenylalanine-histidine.
Particularly preferably, the peptide-based material is diphenylalanine. The inventor of the invention finds that under the preferable condition, the fluorescent sensing material has better selective identification and detection sensitivity to cartap, quicker response and stronger adsorption capacity.
Preferably, the weight ratio of the peptide-based material to the metal ion source is 1: (0.01-2).
More preferably, the weight ratio of the amount of the peptide-based material to the metal ion source is 1: (0.2-1.0). The inventor of the invention finds that under the preferable condition, the fluorescent sensing material has better selective identification and detection sensitivity to cartap, quicker response and stronger adsorption capacity.
Preferably, in the method of manufacturing a light-sensitive peptide element for a fluorescent sensing material, the metal ion in the metal ion source is selected from at least one of zinc ion, nickel ion, copper ion and cobalt ion.
Preferably, the solvent I is used in an amount of 30 to 60mL relative to 100mg of the peptide-based material;
preferably, the solvent I contains (1 to 100) by volume: 1, anhydrous methanol and water.
More preferably, the solvent I contains (5 to 20) by volume: 1, anhydrous methanol and water.
Preferably, the conditions of self-assembly include: the temperature is 85-95 ℃ and the pH value is 4-12.
More preferably, the conditions for self-assembly include: the temperature is 85-95 ℃ and the pH value is 7-11.
According to a preferred embodiment, the method of self-assembling a metal ion source with a peptide-based material in the presence of solvent I comprises:
s1: first mixing part of the solvent I with the metal ion source to obtain a mixture I; and, carrying out second mixing on the rest of the solvent I and the peptide-based material raw materials to obtain a mixture II;
s2: thirdly, mixing the mixture I with the mixture II to obtain a mixture III;
s3: the mixture III is subjected to self-assembly under water bath heating.
The time of the self-assembly is not particularly limited until the anhydrous methanol in the mixture III is evaporated to dryness.
The specific mode of the first mixing, the second mixing and the third mixing is not particularly limited, and may be selected according to technical means known in the art by a person skilled in the art, and is exemplified by mixing for 10 to 40 minutes at an ultrasonic frequency of 30 to 60HZ, which is not repeated herein, and cannot be understood as a limitation of the present invention by a person skilled in the art.
The method for preparing the light-sensitive peptide element for the fluorescent sensing material further comprises conventional post-treatment means in the field such as separation and drying, and the invention provides a preferred specific embodiment by way of example, and the light-sensitive peptide element is obtained by carrying out centrifugal separation on a mixture obtained by evaporating absolute methanol in the mixture III to dryness under the condition of 8000-11000 rpm and then drying the separated precipitate under the condition of 50-70 ℃. The invention is not described in detail herein, and those skilled in the art should not understand the limitation of the invention.
As described above, the third aspect of the present invention provides a light-sensitive peptide element for a fluorescent sensing material prepared by the method of the second aspect.
Preferably, the average particle diameter of the light-sensitive peptide element is 2 to 20nm.
As previously described, a fourth aspect of the present invention provides a method of preparing a fluorescent sensing material, the method comprising: covalently assembling the rigid component with the light-sensitive peptide element in the presence of solvent II and a catalyst; the light-sensitive peptide element is the light-sensitive peptide element for a fluorescent sensing material according to the first or third aspect; the rigid component is at least one selected from 1, 4-benzene dicarboxaldehyde and 4,4' -biphenyl dicarboxaldehyde.
Preferably, the ratio of the light-sensitive peptide element, the rigid component and the catalyst is 1: (0.75-3.75): (1-10).
More preferably, the light-sensitive peptide element, the rigid component and the catalyst are used in a weight ratio of 1: (1.25-2.75): (0.8-5). The inventor of the invention finds that under the preferable condition, the fluorescent sensing material has better selective identification and detection sensitivity to cartap, quicker response and stronger adsorption capacity.
Preferably, the catalyst is acetic acid.
Preferably, the solvent II is selected from at least one of absolute methanol, ethanol and water.
Preferably, the solvent II is used in an amount of 0.50 to 1.25mL relative to 1mg of the light-sensitive peptide element.
Preferably, the conditions of covalent assembly include: the time is 2-48 h, and the temperature is 24-28 ℃.
More preferably, the conditions of covalent assembly include: the time is 2-10 h, and the temperature is 24-28 ℃.
More preferably, the covalent assembly is performed under stirring conditions, the stirring speed being 100-400 rpm.
According to a preferred embodiment, the method further comprises: and sequentially purifying and drying the mixture obtained after the covalent assembly of the light-sensitive peptide element and the rigid component to obtain the fluorescent sensing material.
Preferably, the step of purifying comprises: the mixture obtained after the covalent assembly of the light-sensitive peptide element and the rigid component is mixed with ethanol.
The method for preparing the fluorescent sensing material further comprises conventional post-treatment means in the field such as separation, drying and the like, and the invention provides a preferred specific embodiment by way of example, the mixture obtained after covalent assembly is mixed with ethanol, centrifugal separation is carried out under the condition of 8000-11000 rpm, and then the precipitate obtained after separation is dried under the condition of 50-70 ℃ to obtain the fluorescent sensing material. The invention is not described in detail herein, and those skilled in the art should not understand the limitation of the invention.
As previously described, a fifth aspect of the present invention provides a fluorescent sensing material prepared by the method of the fourth aspect.
Preferably, the average particle diameter of the fluorescent sensing material is 50-100 nm.
As described above, the sixth aspect of the present invention provides the use of the fluorescent sensing material according to the fifth aspect for detecting cartap.
The fluorescent sensing material provided by the invention can detect the residual quantity of cartap in food or other materials.
The fluorescent sensing material provided by the invention also has the following specific advantages:
(1) According to the invention, at least one metal ion selected from zinc ions, nickel ions, copper ions and cobalt ions is self-assembled with at least one peptide-based material selected from diphenyl alanine, carbon end amidated diphenyl alanine, di-histidine and phenylalanine-histidine, in particular diphenyl alanine, so that the peptide-based nano dots are obtained, and the peptide-based nano dots are used as light-sensitive peptide elements, so that the light-sensitive peptide elements have safe and environment-friendly synthetic strategies and have strong optical properties.
(2) The invention covalently connects the peptide-based nano-dots serving as the light-sensitive peptide elements with at least one rigid component selected from 1, 4-benzene dicarboxaldehyde and 4,4' -biphenyl dicarboxaldehyde, and the formed peptide-based nano-dot covalent assembly with a reticular framework structure has the advantages of direct contact sites, cascade response, rapid diffusion capability, safety and environmental protection.
(3) The fluorescent sensing material created by the invention is safe and environment-friendly to synthesize, saves the preparation cost, has excellent detection performance, can rapidly and sensitively detect cartap residues in foods, and provides a basis for later popularization and application.
The invention will be described in detail below by way of examples.
In the examples below, the room temperature was 26.+ -. 2 ℃ unless otherwise indicated.
In the examples below, the various raw materials used were all from commercial sources, unless otherwise specified.
Diphenylalanine: purchased from Shanghai Source leaf Biotechnology Co., ltd. With a purity of 98%.
Carbon-terminal amidated diphenylalanine: purchased from Shanghai Chu peptide biotechnology Co., ltd, purity > 98%.
Di-histidine: purchased from Shanghai Chu peptide biotechnology Co., ltd, purity > 98%.
Phenylalanine-histidine: purchased from Shanghai Chu peptide biotechnology Co., ltd, purity > 98%.
1, 4-benzenedicarboxaldehyde: purchased from Jilin, scientific and technological Co., ltd, and has a purity of 97%.
4,4' -biphenyldicarboxaldehyde: purchased from Jilin, scientific and technological Co., ltd, and has a purity of 97%.
The scanning electron microscope used hereinafter was model SU8020, available from Hitachi corporation.
The type of transmission electron microscope used below was Tecnai G2F 30, available from FEI company.
Preparation example 1
To 50mL centrifuge tubes containing 75mg of diphenylalanine and 35.5mg of zinc chloride, 15mL of solvent I (anhydrous methanol/water, 9:1, v/v) was added, respectively, and the solution was sonicated at room temperature (40 HZ,30 min) to dissolve the same, and then the above 15mL of zinc chloride solution was added to the diphenylalanine solution under sonication conditions, followed by sonication (40 HZ,15 min). Then the obtained mixed solution with the pH value of 10.8 is placed in a water bath environment with the temperature of 85 ℃ until the absolute methanol in the system is completely volatilized. Finally separating the precipitate by a centrifuge under 10000rpm, and drying in a blast oven at 60 ℃ to obtain the light-sensitive peptide element T1.
The result of transmission electron microscopy analysis of the light-sensitive peptide element T1 is shown in FIG. 1. As can be seen from FIG. 1, the average particle diameter of the light-sensitive peptide element T1 is 2 to 20nm, and the light-sensitive peptide element T1 is dispersed as single particles and has a spherical appearance.
Preparation example 2
To 50mL centrifuge tubes containing 75mg of C-terminally amidated diphenylalanine base and 35.5mg of zinc chloride, 15mL of the mixed solvent (anhydrous methanol/water, 9:1, v/v) was added, respectively, and the mixture was dissolved by ultrasonic treatment at room temperature (40 HZ,30 min), and then the above 15mL of zinc chloride solution was added to the C-terminally amidated diphenylalanine base solution under ultrasonic conditions, and the ultrasonic treatment was continued (40 HZ,15 min). Then the obtained mixed solution with the pH value of 10.8 is placed in a water bath environment with the temperature of 85 ℃ until the absolute methanol in the system is completely volatilized. Finally separating the precipitate by a centrifuge, and drying in a blast oven at 60 ℃ to obtain a light-sensitive peptide element T2;
the result of transmission electron microscopy analysis of the light-sensitive peptide element T2 was similar to that of FIG. 1.
Preparation example 3
To 50mL centrifuge tubes containing 75mg of di-histidine and 15mg of cobalt nitrate, 12mL of solvent I (absolute methanol/water, 5:1, v/v) was added, respectively, and the solution was sonicated at room temperature (40 HZ,30 min) to dissolve the same, then the 12mL cobalt nitrate solution was added to the di-histidine solution under sonication conditions, and sonication was continued (40 HZ,15 min). Then the obtained mixed solution with the pH value of 10.8 is placed in a water bath environment with the temperature of 90 ℃ until the absolute methanol in the system is completely volatilized. And finally separating the precipitate by a centrifugal machine, and drying in a blast oven at 60 ℃ to obtain the light-sensitive peptide element T3.
The result of transmission electron microscopy analysis of the light-sensitive peptide element T3 was similar to that of FIG. 1.
Preparation example 4
To 50mL centrifuge tubes containing 75mg phenylalanine-histidine and 35.5mg nickel chloride, 20mL solvent I (absolute methanol/water, 20:1, v/v) was added, respectively, and the solution was sonicated at room temperature (40 HZ,30 min) to dissolve the same, then the above 20mL nickel chloride solution was added to the phenylalanine-histidine solution under sonication conditions, and the sonication was continued (40 HZ,15 min). Then the obtained mixed solution with the pH value of 10.8 is placed in a water bath environment with the temperature of 95 ℃ until the absolute methanol in the system is completely volatilized. And finally separating the precipitate by a centrifugal machine, and drying in a blast oven at 60 ℃ to obtain the light-sensitive peptide element T4.
The result of transmission electron microscopy analysis of the light-sensitive peptide element T4 was similar to that of FIG. 1.
Example 1
4mg of light-sensitive peptide element T1,6mg of 1, 4-benzene dicarboxaldehyde, 2.5mL of absolute methanol, 10. Mu.L of acetic acid (6M) were added to a 25mL round bottom flask, and the mixture was stirred at 26℃and 200rpm for 5 hours. After covalent assembly is finished, the fluorescent sensing material L1 is obtained by purifying the obtained white product with 5mL of ethanol for three times and finally drying the obtained white product in a blast oven at 60 ℃.
The fluorescent sensing material L1 was subjected to scanning electron microscope analysis, and the result is shown in fig. 2. As can be seen from FIG. 2, the fluorescent sensing material is further covalently assembled into a spherical structure having an average particle diameter of 50 to 100nm.
Example 2
4mg of light-sensitive peptide element T2,6mg of 4,4' -biphenyldicarboxaldehyde, 2.5mL of absolute methanol, and 10. Mu.L of acetic acid (6M) were added to a 25mL round bottom flask, and the mixture was stirred at 24℃and 200rpm for 5 hours. After covalent assembly is finished, the mixture is purified by 5mL of ethanol for three times, and finally the obtained white product is placed in a blast oven at 60 ℃ for drying, so as to obtain the fluorescent sensing material L2.
The fluorescent sensing material L2 was subjected to scanning electron microscope analysis, and the result was similar to FIG. 2.
Example 3
4mg of light-sensitive peptide element T3,8mg of 1, 4-benzene dicarboxaldehyde, 2.5mL of ethanol, and 10. Mu.L of acetic acid (6M) were added to a 25mL round bottom flask, and the mixture was magnetically stirred at 28℃and 400rpm for 2 hours. After covalent assembly is finished, the mixture is purified by 5mL of ethanol for three times, and finally the obtained white product is placed in a blast oven at 60 ℃ for drying, so as to obtain the fluorescent sensing material L3.
The fluorescent sensing material L3 was subjected to scanning electron microscope analysis, and the result was similar to FIG. 2.
Example 4
4mg of light-sensitive peptide element T4,5mg of 4,4' -biphenyldicarboxaldehyde, 2.5mL of water, and 10. Mu.L of acetic acid (6M) were added to a 25mL round bottom flask, and the flask was stirred at 26℃and 100rpm for 10 hours. After covalent assembly is finished, the mixture is purified by 5mL of ethanol for three times, and finally the obtained white product is placed in a blast oven at 60 ℃ for drying, so as to obtain the fluorescent sensing material L4.
The fluorescent sensing material L4 was subjected to scanning electron microscope analysis, and the result was similar to FIG. 2.
Test example 1
The light-sensitive peptide element T1 and the fluorescent sensing material L1 prepared in example 1 were respectively subjected to a kinetic response experiment, thereby verifying the cascade response effect and the rapid diffusion capability of the fluorescent sensing material having a network frame structure.
Specifically, absolute methanol is used as a solvent, and cartap standard solution is prepared for standby. 100 mu L of light-sensitive peptide element T1 (1 mg/mL) or 100 mu L of fluorescent sensing material L1 (1 mg/mL) and 100 mu L of cartap standard solution or blank with the concentration of 360 mu g/L are respectively added into a black hole plate with the single hole volume of 400 mu L, uniformly mixed and vibrated for 45min, and the fluorescence intensity of the sample is recorded at fixed time intervals in the process, wherein the fluorescence intensity when the cartap standard solution is added is expressed as F, and the fluorescence intensity when the blank solvent anhydrous methanol is added is expressed as F 0 . After which the time sum F is plotted 0 The response balance time is determined by the relation curve between/F. Wherein time is on the abscissa, F 0 and/F is the ordinate. The instrument parameters were set as follows: gain 115, excitation wavelength was set to 360nm and emission wavelength was set to 570nm.
Fig. 3 and 4 show the fluorescence response law of the light-sensitive peptide element T1 and the fluorescence sensing material L1 to the added cartap at different oscillation times.
As can be seen from FIG. 3, F is present during the contact of the light-sensitive peptide element T1 with cartap for 0-9min 0 The increase in/F over time, in particular, at 2min, shows an opposite trend, since the greater unbalanced concentration of the light-sensitive peptide element T1 when initially contacted with cartap causes a momentary decrease in fluorescence of the light-sensitive peptide element T1. And the fluorescence intensity tends to be smooth with further extension of time.
In contrast, as shown in FIG. 4, F of the fluorescent sensing material L1 0 The F steadily rises and becomes stable after 6min, the reaction time is greatly shortened, the light-sensitive peptide element T1 has uniform and periodic space distribution due to the reticular framework structure obtained by covalent assembly with the rigid component, and more cartap molecules are driven by the reticular framework structure continuously after the cartap molecules are in first contact with the light-sensitive peptide element T1The cartap molecules enter the frame cavity, and then cascade response of the second contact, the third contact and the like is initiated, so that the cartap can be rapidly and efficiently identified.
Test example 2
The adsorption capacity of the fluorescent sensing material L1 prepared in example 1 was measured, thereby verifying the adsorption capacity of the fluorescent sensing material having a mesh-shaped frame structure to cartap.
Specifically, 7 groups of 5mg fluorescent sensing materials L1 are weighed and placed in a 10mL centrifuge tube, 5mL cartap standard solution is added into the centrifuge tube, the concentrations are 10 mg/L, 15 mg/L, 20 mg/L, 25 mg/L, 30 mg/L, 35 mg/L and 40mg/L respectively, the mixture is oscillated for 1h at room temperature, and the supernatant obtained after centrifugation is used for ultraviolet-visible absorption light measurement. Concentration and adsorption capacity (Q) e Mg/g), the maximum adsorption capacity was determined. Wherein the concentration is on the abscissa, adsorption capacity (Q e Mg/g) is on the ordinate. Instrument detection and setting parameters: the sample volume was 900. Mu.L and the detection wavelength was 204nm.
Fig. 5 shows the law of the adsorption amount of the fluorescent sensing material L1 to cartap with different concentrations. As can be seen from fig. 5, in the range of 0-40mg/L, cartap is continuously diffused into the space of the mesh frame cavity of the fluorescent sensing material L1 under the driving of the mesh frame structure as the cartap concentration increases. When the cartap concentration is further increased, the adsorption value of the fluorescent sensing material L1 tends to be stable and does not increase, and the maximum adsorption capacity reaches 217.3mg/L.
The results of testing the light-sensitive peptide elements T2, T3, T4 and the fluorescent sensing materials L2, L3, L4 by the methods similar to those of test example 1 and test example 2 are similar to those of the above-described light-sensitive peptide element T1 and fluorescent sensing material L1, respectively. The fluorescent sensing material prepared by the invention can be used for rapidly detecting cartap residue, and provides a basis for back-end popularization and application.
Test example 3
The fluorescence intensity of the fluorescence sensing materials L1, L2, L3 and L4 prepared in the embodiment is detected.
Specifically, 3mg of each of the fluorescent sensing materials L1, L2, L3, and L4 was accurately weighed, placed in a 5mL centrifuge tube, 1mL of methanol was added thereto, uniformly dispersed by vortexing, 200. Mu.L of each of the above mixtures was sucked up, and added to a 400. Mu.L single-well black well plate, and the fluorescence intensity (a.u.) was recorded. The instrument parameters were set as follows: gain 115, excitation wavelength was set to 390nm, and emission wavelength was set to 570nm.
Fig. 6 shows the fluorescence intensities of the fluorescent sensing materials L1, L2, L3, L4. As can be seen from FIG. 6, the fluorescent sensing materials L1, L2, L3 and L4 have stronger fluorescence intensity, and especially the fluorescent sensing material L1 has stronger fluorescence emission behavior.
Test example 4
Three pesticides are taken and respectively: cartap, fenvalerate and chlorantraniliprole are taken as competitive analogues to verify the adsorption specificity of the fluorescent sensing material L1 to the cartap, so that the high selectivity of the fluorescent sensing material provided by the invention to the cartap is verified.
Specifically, methanol is used as a solvent to prepare cartap, fenvalerate and chlorantraniliprole standard solution with the concentration of 360 mug/L. Then respectively adding 3mg of fluorescent sensing material L1 into 1mL of cartap, fenvalerate, chlorantraniliprole standard solution and methanol, uniformly mixing, vibrating for 45min, and recording the fluorescent intensity of the sample, wherein the fluorescent intensity when the pesticide standard solution is added is represented as F, and the fluorescent intensity when the blank solvent anhydrous methanol is added is represented as F 0 . Thereafter pesticide and F 0 The relation between/F. Wherein the pesticide is on the abscissa, F 0 and/F is the ordinate. The instrument parameters were set as follows: gain 115, excitation wavelength was set to 360nm and emission wavelength was set to 570nm.
Fig. 7 shows the adsorption specificity of the fluorescent sensing material L1 to cartap. As can be seen from FIG. 7, the fluorescent sensing material L1 is used for controlling the F of cartap compared with other pesticides 0 The value of F is 2.20, which is far higher than F for other pesticides 0 and/F, the fluorescent sensing material L1 has better adsorption specificity to cartap. This is due to the effective electron transfer of the fluorescent sensing material L1 and cartap through hydrogen bonding, while other pesticides have stronger electron-donating ability than cartap due to benzene ring or multi-benzene ring structure, which is difficult to realizeEfficient quenching of fluorescence of the fluorescent sensing material L1.
From the analysis, the fluorescent sensing material has the advantages of good selective recognition, high detection sensitivity, quick response and strong adsorption capacity on cartap, and has great practical value and wide application prospect.
The preferred embodiments of the present invention have been described in detail above, but the present invention is not limited thereto. Within the scope of the technical idea of the invention, a number of simple variants of the technical solution of the invention are possible, including combinations of the individual technical features in any other suitable way, which simple variants and combinations should likewise be regarded as being disclosed by the invention, all falling within the scope of protection of the invention.
Claims (10)
1. A light-sensitive peptide element for a fluorescent sensing material, characterized in that the light-sensitive peptide element comprises a peptide-based material and metal particles bridged on the peptide-based material; the peptidyl material is provided by at least one of diphenyl alanine, carbon-terminally amidated diphenyl alanine, di-histidine and phenylalanine-histidine; the metal particles are provided by a metal ion source;
the metal ions in the metal ion source are selected from at least one of zinc ions, nickel ions and cobalt ions.
2. A method of preparing a light-sensitive peptide element for use in a fluorescent sensing material, the method comprising: self-assembling a metal ion source and a peptide-based material in the presence of a solvent I; the peptide-based material is at least one selected from the group consisting of diphenyl alanine, carbon-terminal amidated diphenyl alanine, di-histidine and phenylalanine-histidine; the metal ions in the metal ion source are selected from at least one of zinc ions, nickel ions and cobalt ions;
and/or the weight ratio of the peptide-based material to the metal ion source is 1: (0.01-2).
3. The method of claim 2, wherein the conditions of self-assembly include: the temperature is 50-100 ℃, and the pH value is 4-12.
4. A light-sensitive peptide element for a fluorescent sensing material prepared by the method of claim 2 or 3.
5. The light-sensitive peptide element as claimed in claim 4, wherein the light-sensitive peptide element has an average particle diameter of 2 to 20nm.
6. A method of preparing a fluorescent sensing material, the method comprising: covalently assembling the rigid component with the light-sensitive peptide element in the presence of solvent II and a catalyst; the light-sensitive peptide element is the light-sensitive peptide element for a fluorescent sensing material according to any one of claims 1,4, and 5; the rigid component is at least one selected from 1, 4-benzene dicarboxaldehyde and 4,4' -biphenyl dicarboxaldehyde; the catalyst is acetic acid;
the weight ratio of the light-sensitive peptide element to the rigid component to the catalyst is 1: (0.75-3.75): (1-10).
7. The method of claim 6, wherein the conditions of covalent assembly comprise: the time is 2-48 h, and the temperature is 24-28 ℃.
8. The method according to claim 6 or 7, wherein the method further comprises: and sequentially purifying and drying the mixture obtained after the covalent assembly of the light-sensitive peptide element and the rigid component to obtain the fluorescent sensing material.
9. A fluorescent sensing material prepared by the method of any one of claims 6-8.
10. The use of the fluorescent sensing material of claim 9 for detecting cartap.
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