CN115340615B - Fluorescent molecule based on cyclodextrin-amino acid and synthetic method and application thereof - Google Patents
Fluorescent molecule based on cyclodextrin-amino acid and synthetic method and application thereof Download PDFInfo
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- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
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- C08B37/0009—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid alpha-D-Glucans, e.g. polydextrose, alternan, glycogen; (alpha-1,4)(alpha-1,6)-D-Glucans; (alpha-1,3)(alpha-1,4)-D-Glucans, e.g. isolichenan or nigeran; (alpha-1,4)-D-Glucans; (alpha-1,3)-D-Glucans, e.g. pseudonigeran; Derivatives thereof
- C08B37/0012—Cyclodextrin [CD], e.g. cycle with 6 units (alpha), with 7 units (beta) and with 8 units (gamma), large-ring cyclodextrin or cycloamylose with 9 units or more; Derivatives thereof
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- C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
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- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
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Abstract
The invention relates to a fluorescent molecule based on cyclodextrin-amino acid, a synthesis method and application thereof, wherein the preparation method comprises the following steps: adding aldehyde cyclodextrin and amino acid into water to prepare mixed solution, and sequentially adjusting pH, heating and stirring, and carrying out solid-liquid separation to obtain fluorescent molecules based on cyclodextrin-amino acid; wherein the aldehyde cyclodextrin is aldehyde cyclodextrin obtained by pre-oxidizing beta-cyclodextrin with sodium periodate. Compared with the prior art, the cyclodextrin-histidine fluorescent molecule prepared by the invention can be used as a specific recognition fluorescent probe of aureomycin, has high sensitivity and strong selectivity, and can be used for visual portable detection based on fluorescent images.
Description
Technical Field
The invention belongs to the technical field of fluorescence sensors, and relates to a cyclodextrin-amino acid-based fluorescent molecule, a synthesis method thereof and application thereof in aureomycin fluorescence detection.
Background
Organic fluorescent materials have been widely used in sensors, cell imaging and display technologies. The traditional fluorescent molecules all have the structural basis of chemical bond conjugation based on pi-conjugated aromatic structure, and have adjustable luminescent color and high fluorescent efficiency. However, these materials generally have the characteristics of poor solubility, high biotoxicity, high cost, complex synthesis process and the like, and greatly limit practical application. In contrast, unconventional luminophores without significant conjugated structures have the unique advantages of high biocompatibility, low toxicity, good processibility and ease of synthesis. Unconventional emitters have electron-rich heteroatoms such as nitrogen (N), oxygen (O), sulfur (S), phosphorus (P), halogens (Cl, br and I) or unsaturated groups containing c= O, C =c and c≡n. They emit in concentrated and/or solid state, but tend not to emit light in dilute solutions. This phenomenon is known as Cluster Triggered Emission (CTE). Clustered light emitters in an aggregated state have been used for encryption and bioimaging. However, the concentration of clustered emitters for cell imaging is 1000 times higher than that of conventional emitters, which is a major obstacle to their practical use. Furthermore, in sensing applications, fluorescent probes need to interact effectively with the analyte. However, current solid state clustered light emitting materials based on clustered light emission have limited interactions with analytes in aqueous solutions, and the reported unconventional luminophores have poor performance as sensors. Therefore, there is a great need to find new strategies to promote CTE effects with strong luminescence even in dilute solutions.
The phenomenon of luminescence of mono-, di-, oligo-and polysaccharide crystals having abundant hydroxyl groups as an unconventional luminophore has been widely studied. They are weakly emitted in dilute solutions and bright emissions are only observed from concentrated solutions (> 8 wt.%) or crystalline states. In dilute solutions, linear polysaccharides, such as sodium alginate, exhibit an extended vermiform conformation, which results in non-luminescence due to lack of sufficient electron delocalization and active molecular movement. Thus, the development of nonlinear molecules and enhancement of molecular stacking is of great importance for achieving bright luminescence in dilute solutions.
Compared with the traditional detection method, the fluorescent probe technology is considered to be one of the most promising analysis methods for detecting trace pollutants due to the characteristics of simplicity, portability, high sensitivity, low cost and the like. At present, tetracycline is used as a novel pollutant which is frequently detected in water, and various fluorescent sensing materials have been reported for detecting tetracycline antibiotics. However, it is difficult to distinguish between tetracycline antibiotics such as aureomycin, oxytetracycline, minocycline, tetracycline, etc., which have very similar molecular host structures. Therefore, it is necessary to design a fluorescent probe for a specific tetracycline antibiotic for development and application.
Novel fluorescent molecules based on cyclodextrin-amino acids have not been reported. Reported technologies such as chinese patent CN 201810866243.1 disclose a safe and simple method for preparing double-doped nitrogen and phosphorus carbon quantum dots, which comprises dissolving amino acid and carbon precursor (including glucose, citric acid monohydrate or cyclodextrin) in deionized water, adding phosphoric acid solution, and then performing ultrasonic treatment for 1-2 hours; and heating the solution after ultrasonic treatment in an oil bath at 90-150 ℃ for 1-5 hours to prepare the nitrogen and phosphorus co-doped carbon dot solution. By contrast, although raw materials such as amino acid and cyclodextrin are adopted in the patent, the preparation method and the final product are obviously different, and the nitrogen and phosphorus double-doped carbon quantum dots obtained by adopting methods such as ultrasonic and high-temperature oil bath are long in synthesis time and complex in treatment process. In contrast, the novel fluorescent molecule based on the amino acid grafted cyclodextrin has definite molecular composition and space structure, is different from carbon quantum dots, has mild synthesis conditions, has the synthesis temperature of 60-80 ℃ and has the reaction time of only 30-60min. In addition, the fluorescent molecule of histidine grafted cyclodextrin can be used as a specific recognition probe of aureomycin, can realize high-sensitivity and high-selectivity portable detection of aureomycin, and has good application prospect.
In the aspect of fluorescence analysis technology of aureomycin, a reported technology such as China patent CN201710302455.2 discloses a gold/platinum bimetallic nanocluster fluorescent probe based on polyethyleneimine protection and application thereof in aureomycin detection, and the method utilizes the polyethyleneimine-protected bimetallic gold/platinum nanocluster as a fluorescence sensing material and is used for detecting tetracycline antibiotics based on a fluorescence quenching mechanism; however, it is still necessary to add Al 3+ Based on fluorescence enhancement mechanism, aureomycin can be distinguished from tetracycline, so that the purpose of specifically detecting aureomycin is achieved. The method is used for detecting aureomycin, the linear range is 0.5-30 mu M, and the detection limit is 0.5 mu M. However, the detection limit of the detection method is still high, and the detection requirement of trace aureomycin in the existing sewage is difficult to meet.
Disclosure of Invention
The invention aims to provide a fluorescent molecule based on cyclodextrin-amino acid, and a synthesis method and application thereof.
The aim of the invention can be achieved by the following technical scheme:
a method for synthesizing a cyclodextrin-amino acid based fluorescent molecule, comprising:
adding aldehyde cyclodextrin and amino acid into water to prepare mixed solution, and sequentially adjusting pH, heating and stirring, and carrying out solid-liquid separation to obtain fluorescent molecules based on cyclodextrin-amino acid; wherein the aldehyde cyclodextrin is aldehyde cyclodextrin obtained by pre-oxidizing beta-cyclodextrin by sodium periodate.
Further, the preparation method of the aldehyde cyclodextrin comprises the following steps:
stirring beta-cyclodextrin and sodium periodate in water in a dark place for reaction, stirring and mixing the mixture with ethanol after nanofiltration until precipitation is achieved, and then filtering, washing and freeze-drying the mixture in sequence to obtain aldehyde cyclodextrin;
wherein, the mol ratio of the beta-cyclodextrin to the sodium periodate is 1 (1-4);
in the light-shielding stirring reaction, the reaction temperature is 30-50 ℃ and the reaction time is 3-5h;
the purification process adopts a water/ethanol mixed solution with the volume ratio of 1/4 for washing.
Further, the molar ratio of the aldehyde cyclodextrin to the amino acid is 1 (2-6).
Further, the amino acid includes at least one of glycine, isoleucine, methionine, cysteine, glutamic acid, glutamine, asparagine, arginine, lysine, phenylalanine, tryptophan or histidine.
Further, when the amino acid is at least one of glycine, isoleucine, methionine, cysteine, glutamic acid, glutamine, asparagine, arginine, lysine or phenylalanine, the pH of the reaction system is adjusted to 8-9;
when the amino acid is one or two of tryptophan and histidine, the pH of the reaction system is adjusted to 6-7.
Further, in the heating and stirring process, the heating temperature is 60-80 ℃, and the stirring time is 30-60min.
Further, the solid-liquid separation comprises dialysis and freeze drying.
Fluorescent molecules based on cyclodextrin-amino acids are synthesized using the methods described above.
The application of the cyclodextrin-amino acid-based fluorescent molecule comprises the step of taking the fluorescent molecule as a fluorescent probe for qualitative and/or quantitative detection of aureomycin in a water body.
Further, the detection method specifically includes the following steps:
1) Drawing a standard curve: respectively mixing and uniformly stirring the fluorescent probes with a plurality of solutions containing aureomycin with different concentrations to obtain a standard solution with aureomycin concentration range of 0-5 mu M, taking a 365nm ultraviolet lamp as an excitation light source, shooting in a dark environment to obtain a fluorescent image, analyzing the B value and the G value of the image, drawing a standard curve with B/G as an ordinate and aureomycin concentration as an abscissa, and detecting and fitting an equation of aureomycin;
2) Detecting aureomycin in a water sample: mixing the fluorescent probe with a water sample to be detected by adopting the dosage ratio of the fluorescent probe to the solution of the aureomycin in the step 1) to obtain a mixed sample, obtaining a fluorescent image by taking a 365nm ultraviolet lamp as an excitation light source, analyzing and calculating to obtain B/G of the image, and obtaining the corresponding aureomycin concentration according to a standard curve or a fitting equation.
Firstly, carrying out oxidation modification on beta-cyclodextrin with low solubility and low reactivity by utilizing sodium periodate to obtain aldehyde cyclodextrin with good water solubility and high reactivity and containing a dialdehyde structure; then, the aldehyde group of the aldehyde cyclodextrin reacts with amino groups in various amino acid molecules through Schiff base to obtain novel 12 amino acid grafted cyclodextrin fluorescent molecules based on different types of amino acids. The synthesized fluorescent molecules have excellent fluorescent properties in dilute solutions based on the mechanism of enhanced clustered light emission, good water solubility and excellent environmental compatibility. The synthesized histidine grafted cyclodextrin fluorescent molecule is used as a fluorescent probe of aureomycin, and can realize high-selectivity and high-sensitivity portable detection of aureomycin based on a visualization technology.
Compared with the prior art, the invention has the following characteristics:
1) The synthesis method is simple, the cyclodextrin is obtained after being modified simply, the synthesis condition is mild, the reaction temperature can be controlled below 80 ℃, and the preparation environment requirement can be provided based on the conventional water bath condition;
2) Compared with the traditional fluorescent molecule with a conjugated structure, the novel fluorescent molecule of the cyclodextrin-amino acid has the advantages of simple structure, low synthesis cost, good water solubility, no biotoxicity, good environmental compatibility and the like.
3) The invention can synthesize a series of cyclodextrin-amino acid novel fluorescent molecules with rich structures and performances, and provides a general method for synthesizing an unconventional fluorescent material based on amino acid-cyclodextrin.
4) The novel fluorescent molecule of cyclodextrin-histidine prepared by the invention can directly identify aureomycin molecules with high characteristics, realizes high-sensitivity detection (detection range is 0-5 mu M, detection limit is 12 nM) of aureomycin, is more suitable for accurate detection of low-concentration trace pollutants and high-selectivity detection, has no signal response to other tetracyclines with very similar molecular main structures, and can realize portable detection of aureomycin by a visualization technology based on fluorescent images.
Drawings
FIG. 1 is a three-dimensional fluorescence of fluorescent molecules of cyclodextrin-amino acids synthesized in example 1 and example 2;
FIG. 2 is a graph showing the emission spectra of fluorescent molecules of cyclodextrin-amino acids synthesized in example 1 and example 2 under different excitation wavelengths;
FIG. 3 is a fluorescence spectrum of cyclodextrin-histidine at various amino acid additions in example 3;
FIG. 4 is a fluorescence emission spectrum of example 4 after adding chlortetracycline at various concentrations;
FIG. 5 is a standard curve of aureomycin assay plotted in example 4;
FIG. 6 is the effect of different small organic molecules on the fluorescence emission spectrum of the synthesized cyclodextrin-histidine fluorescent molecule in example 5.
Detailed Description
The invention will now be described in detail with reference to the drawings and specific examples.
A cyclodextrin-amino acid based fluorescent molecule, the synthesis method comprising the steps of:
s1: mixing beta-cyclodextrin and sodium periodate in a molar ratio of 1 (1-4) in water, stirring and reacting for 3-5 hours at 30-50 ℃ in a dark place, stirring and mixing the mixture with ethanol after nanofiltration until precipitation is achieved, and then filtering, purifying and freeze-drying the mixture in sequence to obtain aldehyde cyclodextrin;
wherein, the purification process is to wash with water/ethanol mixed solution with volume ratio of 1/4;
s2: preparing aldehyde cyclodextrin into aqueous solution;
s3: adding amino acid into the aqueous solution, regulating the pH value of the system, heating to 60-80 ℃, and continuously stirring for 30-60min to obtain a product liquid;
wherein, the molar ratio of aldehyde cyclodextrin to amino acid is 1 (2-6);
when the amino acid is at least one of glycine, isoleucine, methionine, cysteine, glutamic acid, glutamine, asparagine, arginine, lysine or phenylalanine, regulating the pH of the reaction system to 8-9;
when the amino acid is one or two of tryptophan and histidine, regulating the pH value of the reaction system to 6-7;
s4: dialyzing the product liquid for 12 hours, and freeze-drying the obtained dialysate to obtain the novel fluorescent material powder based on cyclodextrin-amino acid.
The application of the cyclodextrin-amino acid-based fluorescent molecule comprises the step of taking the fluorescent molecule as a fluorescent probe for qualitative and/or quantitative detection of aureomycin in a water body.
Further, the detection method specifically includes the following steps:
1) Drawing a standard curve: respectively mixing and uniformly stirring the fluorescent probes with a plurality of solutions containing aureomycin with different concentrations to obtain a standard solution with aureomycin concentration range of 0-5 mu M, taking a 365nm ultraviolet lamp as an excitation light source, shooting in a dark environment to obtain a fluorescent image, performing image color homogenization treatment by using a software image J, directly reading the average B value and the G value of the image by using an F color sampler, drawing a standard curve with B/G as an ordinate and aureomycin concentration as an abscissa, and detecting and fitting an equation of aureomycin;
2) Detecting aureomycin in a water sample: mixing the fluorescent probe with a water sample to be detected by adopting the dosage ratio of the fluorescent probe to the solution of the aureomycin in the step 1) to obtain a mixed sample, obtaining a fluorescent image by taking a 365nm ultraviolet lamp as an excitation light source, analyzing and calculating to obtain B/G of the image, and obtaining the corresponding aureomycin concentration according to a standard curve or a fitting equation.
The present embodiment is implemented on the premise of the technical scheme of the present invention, and a detailed implementation manner and a specific operation process are given, but the protection scope of the present invention is not limited to the following examples.
Example 1:
a cyclodextrin-amino acid (histidine/tryptophan) -based fluorescent molecule, the synthesis method comprising the steps of:
s1: adding 15g of beta-cyclodextrin into 100mL of deionized water, uniformly stirring, adding 6g of sodium periodate, stirring at 40 ℃ in a dark place for reaction for 4 hours, filtering by a 220nm filter membrane, taking a filtrate, mixing the filtrate with excessive absolute ethyl alcohol (800 mL) until precipitation is separated out, and sequentially filtering, washing for multiple times by ethanol/water (V/V=80/20), and freeze-drying to obtain aldehyde-based cyclodextrin which has good water solubility and high reactivity and contains a dialdehyde structure;
s2: 0.1mmol (114 mg) of aldehyde cyclodextrin and 0.6mmol (93 mg) of histidine or 0.6mmol (82 mg) of tryptophan are dissolved in 20mL of water together, and the pH of the system is adjusted to 6-7;
s3: stirring the solution at 80 ℃ for reaction for 60min to obtain cyclodextrin-amino acid fluorescent molecules, dialyzing (molecular weight cut-off 1000 Da), and freeze-drying to obtain 2 kinds of fluorescent material powder based on cyclodextrin-amino acid (histidine/tryptophan) respectively.
Example 2:
a fluorescent molecule based on cyclodextrin-amino acids (glycine, isoleucine, methionine, cysteine, glutamic acid, glutamine, asparagine, arginine, lysine or phenylalanine) which differs from example 1 only in the way in which it is synthesized:
in step S2, the amino acid used was 0.6mmol (45 mg) glycine or 0.6mmol (79 mg) isoleucine, 0.6mmol (105 mg) arginine, 0.6mmol (79 mg) asparagine, 0.6mmol (73 mg) cysteine, 0.6mmol (88 mg) glutamine, 0.6mmol (88 mg) glutamic acid, 0.6mmol (88 mg) lysine, 0.6mmol (90 mg) methionine, 0.6mmol (99 mg) phenylalanine; regulating the pH value of the system to 8-9;
in the step S3, stirring and reacting for 30min to finally obtain 10 fluorescent material powders based on cyclodextrin-amino acid;
the procedure is as in example 1.
As shown in FIG. 1, three-dimensional fluorescence of 12 novel fluorescent molecules (5 mM aqueous solution) based on cyclodextrin-amino acids synthesized in example 1 and example 2 was shown. It can be seen from the figure that the novel fluorescent molecules of the synthesized cyclodextrin-amino acids have a distinct fluorescent signal, although without pi-conjugated groups, and the emission wavelength is concentrated in the range of 350-550nm, based on the enhanced clustered light-emitting effect in the cyclodextrin-limited space.
As shown in FIG. 2, the emission spectra of 12 novel fluorescent molecules (5 mM aqueous solution) based on cyclodextrin-amino acids were synthesized at different excitation wavelengths. It can be seen from the graph that the synthesized novel fluorescent molecules have the dependence of excitation wavelength, and the emission peak gradually red-shifts with the increase of the excitation wavelength.
Example 3: influence of the addition of different amino acids
A cyclodextrin-amino acid (histidine) -based fluorescent molecule, the synthesis of which differs from that of example 1 only in that:
in the step S2, the amount of histidine used is 31mg,62mg,93mg and 124mg respectively (molar ratio of aldehyde cyclodextrin to amino acid is 1:2,1:4,1:6 and 1:8 respectively); the procedure is as in example 1.
As a result of measuring the fluorescence emission curve at a concentration of 5mM (aqueous solution) using a 365nm ultraviolet lamp as an excitation light source, the fluorescence intensity gradually increased with increasing histidine addition amount as shown in FIG. 3. When the molar ratio is 1:8, the increase in fluorescence intensity is not significant.
Example 4:
in this example, the cyclodextrin-histidine-based fluorescent molecule prepared in example 1 is used as a fluorescent probe for detecting aureomycin in a water sample, and the specific detection process is as follows:
1) Drawing a standard curve: dispersing cyclodextrin-amino acid-based fluorescent molecules into deionized water at a concentration of 200 μm for use as a fluorescent probe;
respectively preparing chlortetracycline standard solutions with the concentrations of 0, 1, 2, 4, 6, 8 and 10 mu M, respectively taking 0.5mL of the chlortetracycline standard solutions and mixing with a fluorescent probe with the same volume to obtain mixed solution, measuring the fluorescence emission curve of the obtained mixed solution by using a 365nm ultraviolet lamp as an excitation light source in a dark environment, and as shown in the result, as shown in figure 4, the fluorescence intensity of the system is gradually enhanced along with the increase of the concentration of the added chlortetracycline;
photographing the mixed solution to obtain a fluorescence image, gradually increasing the brightness of the image along with the increase of the aureomycin concentration, analyzing the B value and the G value of the image by using image analysis software, drawing a standard curve (shown in fig. 5) by taking B/G as an ordinate and taking aureomycin concentration (mu M) as an abscissa, and obtaining an aureomycin detection fitting equation: y=0.1431x+1.3319, (r2=0.998); the detection limit was calculated from lod=3σ/N (where σ is the standard deviation of the blank sample and N is the slope of the linear equation), and the detection limit was 12nM with the fluorescent molecule of cyclodextrin-histidine as the probe.
2) Pretreatment of a water sample to be tested: filtering a water sample to be detected, and regulating the pH value of the water sample to be detected to be neutral;
3) Detecting aureomycin in a water sample: taking 0.5mL of pretreatment water sample, mixing with 0.5mL of fluorescent probe solution to obtain a mixed sample, obtaining a fluorescent image by taking a 365nm ultraviolet lamp as an excitation light source, obtaining B/G of the image by using image analysis software, and obtaining the corresponding aureomycin concentration according to a fitting equation. The college (four-level road school area) campus water and tap water were collected for standard addition recovery experiments, 1. Mu.M, 3. Mu.M, 4. Mu.M and 7. Mu.M of aureomycin were added respectively, and the above method was used for testing, and the recovery rate of aureomycin was 97.25% -101.67%, and the specific results are shown in Table 1.
Table 1 labeling recovery experiment results
Example 5:
the cyclodextrin-histidine-based fluorescent probe synthesized in example 1 is used for selectively detecting different coexisting or structurally similar small molecular organic matters, and the specific method is as follows:
dispersing cyclodextrin-amino acid-based fluorescent molecules into deionized water at a concentration of 200 μm for use as a fluorescent probe; meanwhile, a plurality of 200 mu M of different small molecule organic matter solutions are respectively prepared and comprise aureomycin (CTC), acetic Acid (AC) Glucose (Glucose), ampicillin (Amp), chloramphenicol (CAP), streptomycin (Str), nalidixic acid (Nal), glycine (Gly), histidine (His), phenylalanine (Phe), tryptophan (Trp), trichloroacetamide (TCAM), tetracycline (TC), terramycin (OTC) and Minocycline (MOC), 1mL of each solution is mixed with an equal volume of fluorescent probe solution, after the reaction system is stable, the fluorescent spectrum is tested (an ultraviolet lamp with the wavelength of 365nm is used as an excitation light source) to examine the selective recognition capability, as shown in figure 6, the fluorescent signal of the system is obviously enhanced after the aureomycin is added, and the emission peak is blue-shifted, and the fluorescent response of the system after other small molecule organic matters are added is not obviously changed, so that the method has good selectivity and the specific recognition of the cyclodextrin-histidine fluorescent probe on the aureomycin is realized.
The previous description of the embodiments is provided to facilitate a person of ordinary skill in the art in order to make and use the present invention. It will be apparent to those skilled in the art that various modifications can be readily made to these embodiments and the generic principles described herein may be applied to other embodiments without the use of the inventive faculty. Therefore, the present invention is not limited to the above-described embodiments, and those skilled in the art, based on the present disclosure, should make improvements and modifications without departing from the scope of the present invention.
Claims (8)
1. A method for synthesizing a fluorescent molecule based on cyclodextrin-amino acid, comprising:
adding aldehyde cyclodextrin and amino acid into water to prepare mixed solution, and sequentially adjusting pH, heating and stirring, and carrying out solid-liquid separation to obtain fluorescent molecules based on cyclodextrin-amino acid; wherein the aldehyde cyclodextrin is aldehyde cyclodextrin obtained by pre-oxidizing beta-cyclodextrin by sodium periodate;
the preparation method of the aldehyde cyclodextrin comprises the following steps:
stirring beta-cyclodextrin and sodium periodate in water in a dark place for reaction, stirring and mixing the mixture with ethanol after nanofiltration until precipitation is achieved, and then filtering, washing and freeze-drying the mixture in sequence to obtain aldehyde cyclodextrin;
wherein, the mol ratio of the beta-cyclodextrin to the sodium periodate is 1 (1-4);
in the light-shielding stirring reaction, the reaction temperature is 30-50 ℃ and the reaction time is 3-5h;
the molar ratio of the aldehyde cyclodextrin to the amino acid is 1 (2-6).
2. The method of claim 1, wherein the amino acid comprises at least one of glycine, isoleucine, methionine, cysteine, glutamic acid, glutamine, asparagine, arginine, lysine, phenylalanine, tryptophan, or histidine.
3. The method for synthesizing a cyclodextrin-amino acid-based fluorescent molecule according to claim 2, wherein when the amino acid is at least one of glycine, isoleucine, methionine, cysteine, glutamic acid, glutamine, asparagine, arginine, lysine or phenylalanine, the pH of the reaction system is adjusted to 8-9;
when the amino acid is one or two of tryptophan and histidine, the pH of the reaction system is adjusted to 6-7.
4. The method for synthesizing fluorescent molecules based on cyclodextrin-amino acid according to claim 1, wherein the heating temperature is 60-80 ℃ and the stirring time is 30-60min during the heating and stirring process.
5. The method for synthesizing a fluorescent molecule based on cyclodextrin-amino acids according to claim 1, wherein the solid-liquid separation comprises dialysis and freeze-drying.
6. A cyclodextrin-amino acid based fluorescent molecule synthesized by the method of any one of claims 1 to 5.
7. The use of a fluorescent molecule based on cyclodextrin-amino acids according to claim 6, wherein said fluorescent molecule is used as a fluorescent probe for qualitative and/or quantitative detection of aureomycin in a body of water.
8. The use of a cyclodextrin-amino acid based fluorescent molecule according to claim 7, wherein the detection method comprises the steps of:
1) Drawing a standard curve: respectively mixing and uniformly stirring the fluorescent probes with a plurality of solutions containing aureomycin with different concentrations to obtain a standard solution with aureomycin concentration range of 0-5 mu M, taking a 365nm ultraviolet lamp as an excitation light source, shooting in a dark environment to obtain a fluorescent image, analyzing the B value and the G value of the image, drawing a standard curve with B/G as an ordinate and aureomycin concentration as an abscissa, and detecting and fitting an equation of aureomycin;
2) Detecting aureomycin in a water sample: mixing the fluorescent probe with a water sample to be detected by adopting the dosage ratio of the fluorescent probe to the solution of the aureomycin in the step 1) to obtain a mixed sample, obtaining a fluorescent image by taking a 365nm ultraviolet lamp as an excitation light source, analyzing and calculating to obtain B/G of the image, and obtaining the corresponding aureomycin concentration according to a standard curve or a fitting equation.
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| CN104383554A (en) * | 2002-09-06 | 2015-03-04 | 天蓝制药公司 | Cyclodextrin-based polymers for therapeutics delivery |
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| CN104383554A (en) * | 2002-09-06 | 2015-03-04 | 天蓝制药公司 | Cyclodextrin-based polymers for therapeutics delivery |
| CN103052387A (en) * | 2010-07-22 | 2013-04-17 | 德国麦氏大药厂 | Pharmaceutical composition containing a tryptophan derivative |
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