CN114682789A - Preparation method of chicken protein protection gold platinum nanocluster and application of chicken protein protection gold platinum nanocluster in methotrexate detection - Google Patents
Preparation method of chicken protein protection gold platinum nanocluster and application of chicken protein protection gold platinum nanocluster in methotrexate detection Download PDFInfo
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- CN114682789A CN114682789A CN202210312460.2A CN202210312460A CN114682789A CN 114682789 A CN114682789 A CN 114682789A CN 202210312460 A CN202210312460 A CN 202210312460A CN 114682789 A CN114682789 A CN 114682789A
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B22—CASTING; POWDER METALLURGY
- B22F—WORKING METALLIC POWDER; MANUFACTURE OF ARTICLES FROM METALLIC POWDER; MAKING METALLIC POWDER; APPARATUS OR DEVICES SPECIALLY ADAPTED FOR METALLIC POWDER
- B22F9/00—Making metallic powder or suspensions thereof
- B22F9/16—Making metallic powder or suspensions thereof using chemical processes
- B22F9/18—Making metallic powder or suspensions thereof using chemical processes with reduction of metal compounds
- B22F9/24—Making metallic powder or suspensions thereof using chemical processes with reduction of metal compounds starting from liquid metal compounds, e.g. solutions
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y40/00—Manufacture or treatment of nanostructures
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- 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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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
- G01N2021/6439—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" with indicators, stains, dyes, tags, labels, marks
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- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
Abstract
The invention relates to the technical field of gold-platinum nanocluster synthesis, in particular to a preparation method of a chicken protein protection gold-platinum nanocluster and application of the chicken protein protection gold-platinum nanocluster in methotrexate detection, and the method comprises the following specific steps: s1, weighing a certain amount of egg white, adding 100 mu L of water for dissolving, adding chloroauric acid solution for mixing uniformly, adding chloroplatinic acid solution, and stirring and mixing uniformly for 2 minutes to obtain solution A; s2, adding a 1M sodium hydroxide solution into the solution A obtained in the step S1, adjusting the pH of the solution to 9-13, and heating the mixed solution in a water bath for 3-10 hours to obtain a solution B; s3, storing the solution B obtained in the step 2 at 4 ℃ to obtain the chicken protein-gold platinum nanocluster. The invention takes the chicken protein as a template, adopts a one-step synthesis method to prepare the chicken protein-gold platinum nanocluster in a water solution, and has unique photophysical characteristics, simple preparation, low toxicity and high stability; the method for detecting the methotrexate is quick and simple, has high detection sensitivity, and is an ideal fluorescent nano material applied to the fields of biology and medicine.
Description
Technical Field
The invention relates to the technical field of gold-platinum nanocluster synthesis, in particular to a preparation method of a chicken protein protection gold-platinum nanocluster and application of the chicken protein protection gold-platinum nanocluster in methotrexate detection.
Background
The egg white is also called as egg white, is transparent high-viscosity liquid surrounding the egg yolk in the eggs, is rich in nutrition, and can replenish vital essence and energy, moisten lung and relieve sore throat, and enhance the immune function of skin. About 90% of the solids of chicken protein is protein, and egg white protein is the major component of protein. Ovalbumin is a glycoprotein containing a sufficient number of amino acid residues, of which cysteine and histidine residues are the protecting agents for metal ions; tyrosine residues can be used as reducing agents to reduce noble metal ions under alkaline conditions. In addition, the chicken protein has low price, easily obtained materials, green and economic properties, and is an ideal material for synthesizing metal nano clusters.
The metal nanoclusters are generally composed of several to several tens of atoms, have a size of a sub-nanometer level, have excellent photostability, a large stokes shift and low biotoxicity, and exhibit strong light absorption through electron transitions between molecular energy levels, thereby generating fluorescence. In addition, the fluorescent material has good selectivity, is an ideal fluorescent sensor, and has important potential in the aspects of catalysis, biomolecule and heavy metal detection, biosensors and the like as a novel nano material. The protein has a plurality of active sites, can be chelated with metal, is often used as a reducing agent and a stabilizing agent for synthesizing the metal nanocluster, has excellent biocompatibility and low toxicity, and is a common ligand for synthesizing the metal nanocluster.
Methotrexate is an antineoplastic drug, is also suitable for rheumatoid arthritis patients who have no effect on the treatment of non-steroidal anti-inflammatory drugs, and can be used for treating osteosarcoma and lymphoma of adults and children at higher doses. Methotrexate is a cytotoxic compound, and has effects on not only tumor cells but also normal cells and tissues, and gastrointestinal damage, hepatorenal toxicity and the like are easily caused by long-term use, so that timely monitoring is required. Aiming at the limitations of high detection cost, low sensitivity, long time consumption and the like of the traditional detection method of methotrexate, the development of a quick and efficient method for accurately detecting the methotrexate in a biological sample is urgently needed.
Disclosure of Invention
The invention aims to solve the defects in the prior art, and provides a preparation method of a chicken protein protection gold platinum nano-cluster and application of the chicken protein protection gold platinum nano-cluster in methotrexate detection.
In order to achieve the purpose, the invention adopts the following technical scheme:
a preparation method of an albumin-protected gold-platinum nano-cluster comprises the following specific steps:
s1, weighing a certain amount of egg white, adding 100 mu L of water for dissolving, adding chloroauric acid solution for mixing uniformly, adding chloroplatinic acid solution, and stirring and mixing uniformly for 2 minutes to obtain solution A;
s2, adding a 1M sodium hydroxide solution into the solution A obtained in the step S1, adjusting the pH of the solution to 9-13, and heating the mixed solution in a water bath for 3-10 hours to obtain a solution B;
s3, storing the solution B obtained in the step 2 at 4 ℃ to obtain the chicken protein-gold platinum nanocluster.
Preferably, in S1, the concentration ratio of chloroauric acid to chloroplatinic acid is 6:1, and the concentration of chicken protein is 50mgml-1。
Preferably, in the S2, the pH value of the solution is adjusted to 13 by sodium hydroxide, the heating temperature is 55 ℃, and the reaction time is 10 hours.
The invention also provides application of the chicken protein-gold platinum nanocluster obtained by the preparation method of the chicken protein protection gold platinum nanocluster in methotrexate detection, the chicken protein-gold platinum nanocluster is diluted by phosphate buffer solution, methotrexate with different concentrations is added and uniformly mixed, the mixture is incubated at room temperature, and under the condition of excitation wavelength, the fluorescence intensity of the chicken protein-gold platinum nanocluster is gradually weakened along with the gradual increase of the concentration of the methotrexate, so that the detection is realized.
Preferably, the detection is achieved under 360nm excitation wavelength conditions with incubation for 10 minutes at room temperature.
Compared with the prior art, the invention has the following beneficial effects:
1. the invention takes the chicken protein as a template, adopts a one-step synthesis method to prepare the chicken protein-gold platinum nanocluster in an aqueous solution, and has unique photophysical characteristics, low toxicity, good stability and excellent biocompatibility.
2. The chicken protein-gold platinum nanocluster prepared by the invention is rapid and simple in methotrexate detection method and good in detection sensitivity, and is an ideal fluorescent nanomaterial applied to the fields of biology and medicine.
Drawings
FIG. 1 shows excitation and emission spectra of the chicken protein-gold platinum nanoclusters of the present invention;
FIG. 2 is a fluorescence spectrum of chicken protein-Au-Pt nanoclusters synthesized by different Au-Pt ratios according to the present invention;
FIG. 3 is a fluorescence spectrum of the chicken protein-gold platinum nanoclusters synthesized by different pH values according to the present invention;
FIG. 4 is a fluorescence spectrum of chicken protein-Au-Pt nanoclusters synthesized by different reaction times according to the present invention;
FIG. 5 is a fluorescence spectrum of a chicken protein-Au-Pt nanocluster synthesized by different chicken protein concentrations according to the present invention;
FIG. 6 is a fluorescence spectrum of chicken protein-gold platinum nanoclusters synthesized at different temperatures according to the present invention;
FIG. 7 is a graph of fluorescence emission spectra of the chicken protein-gold platinum nanoclusters after adding methotrexate of various concentrations in accordance with the present invention;
FIG. 8 is a graph of the linear relationship between fluorescence intensity of methotrexate and the CEPT nanoclusters at different concentrations of the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings, so that those skilled in the art can better understand the advantages and features of the present invention, and thus the scope of the present invention is more clearly defined. The embodiments described herein are only a few embodiments of the present invention, rather than all embodiments, and all other embodiments that can be derived by one of ordinary skill in the art without inventive faculty based on the embodiments described herein are intended to fall within the scope of the present invention.
Referring to fig. 1 to 8, in order to improve the luminescence property of the product, the following steps are included:
weighing a certain amount of egg white, adding 100 mu L of water for dissolving, adding 100 mu L of chloroauric acid solution for uniformly mixing, adding 50 mu L of chloroplatinic acid solution for uniformly mixing, then violently stirring for 2 minutes, adding 1M sodium hydroxide solution into the solution, uniformly mixing to adjust the pH value of the solution, and heating in a water bath for a period of time; and detecting the excitation spectrum and the emission spectrum of the product by using a fluorescence spectrometer. Changing the volume of the chloroplatinic acid to adjust the concentration ratio of the chloroauric acid to the chloroplatinic acid, and observing a fluorescence spectrum, wherein when the concentration ratio of the chloroauric acid to the chloroplatinic acid is 6:1, the fluorescence intensity is strongest; therefore, the concentration ratio of chloroauric acid to chloroplatinic acid is selected to be 6:1 as the optimal ratio for preparing the chicken protein-gold platinum nanoclusters.
Weighing a certain amount of egg white, adding 100 mu L of water for dissolving, adding 100 mu L of chloroauric acid solution for uniformly mixing, adding 16.7 mu L of chloroplatinic acid solution for uniformly mixing, violently stirring for 2 minutes, adding 13 mu L of sodium hydroxide solution with the concentration of 1M into the solution, uniformly mixing, adjusting the pH value of the solution to 9, and heating in a water bath for a period of time; and detecting the excitation spectrum and the emission spectrum of the product by using a fluorescence spectrometer. Increasing the volume of the sodium hydroxide to change the pH value of the solution, observing the fluorescence spectra at different pH values, gradually increasing the fluorescence intensity along with the increase of the pH value of the solution, and when the pH value of the solution is 13, the fluorescence intensity reaches the strongest value; thus pH 13 was chosen as the optimum pH for the preparation of the chicken protein-gold platinum nanoclusters.
Weighing a certain amount of egg white, adding 100 mu L of water for dissolving, then adding 100 mu L of chloroauric acid solution for uniformly mixing, then adding 16.7 mu L of chloroplatinic acid solution for uniformly mixing, then violently stirring for 2 minutes, adding 27 mu L of sodium hydroxide solution with the concentration of 1M into the solution, uniformly mixing, adjusting the pH value of the solution to 13, and heating in a water bath for 3 hours; the emission spectrum of the product was detected using a fluorescence spectrometer. Increasing the reaction time, observing the fluorescence spectra of different reaction times, and gradually increasing the fluorescence intensity along with the increase of the reaction time, wherein the fluorescence intensity reaches the strongest intensity in 10 hours of the reaction time; thus 10 hours was chosen as the optimal reaction time for the preparation of the chicken protein-gold platinum nanoclusters.
Weighing 1mg of chicken protein, adding 100 mu L of water for dissolving, adding 100 mu L of chloroauric acid solution for uniformly mixing, adding 16.7 mu L of chloroplatinic acid solution for uniformly mixing, violently stirring for 2 minutes, adding 27 mu L of sodium hydroxide solution with the concentration of 1M into the solution, uniformly mixing, adjusting the pH value of the solution to 13, and heating in a water bath for 10 hours; the emission spectrum of the product was detected using a fluorescence spectrometer. Increasing the concentration of chicken protein, observing fluorescence spectrum, increasing the fluorescence intensity with the increase of chicken protein concentration, and using 50mg ml-1The fluorescence intensity is strongest when the chicken protein is prepared; thus 50mg ml was selected-1The chicken protein is used as the optimal concentration for preparing the chicken protein-gold platinum nano cluster.
Weighing 5mg of chicken protein, adding 100 mu L of water for dissolving, adding 100 mu L of chloroauric acid solution for uniformly mixing, then adding 16.7 mu L of chloroplatinic acid solution for vigorously stirring for 2 minutes, adding 27 mu L of 1M sodium hydroxide solution into the solution, uniformly mixing, adjusting the pH value of the solution to 13, and heating in a water bath at 20 ℃ for 10 hours; the emission spectrum of the product was detected using a fluorescence spectrometer. Increasing the heating temperature of the water bath, observing the fluorescence spectrum, gradually increasing the fluorescence intensity along with the increase of the water bath temperature, and when the reaction temperature is 55 ℃, the fluorescence intensity is optimal; thus 55 ℃ was chosen as the optimal temperature for the preparation of the chicken protein-gold platinum nanoclusters.
Detection of methotrexate by using chicken protein-gold platinum nanocluster as fluorescent probe
Diluting the chicken protein-gold platinum nanoclusters by using a phosphate buffer solution, adding a certain amount of methotrexate, incubating the mixed solution at room temperature for 10 minutes, and gradually reducing the fluorescence intensity of the chicken protein-gold platinum nanoclusters along with the gradual increase of the concentration of the methotrexate under the condition of an excitation wavelength of 360 nm.
Preparation and optimization of (I) chicken protein-gold platinum nanocluster
The first embodiment is as follows: weighing 5mg of chicken protein, and adding 100 mu L of water for dissolving; adding 100 mu L of chloroauric acid, mixing uniformly, adding 16.7 mu L of chloroplatinic acid solution, mixing uniformly and stirring for 2 minutes; and secondly, adding 27 mu L of 1M sodium hydroxide solution into the solution, uniformly mixing, adjusting the pH value of the solution to 13, and heating in a water bath at 55 ℃ for 10 hours to obtain the chicken protein-gold platinum nanocluster.
Example two: weighing 5mg of chicken protein, and adding 100 mu L of water for dissolving; adding 100 mu L of chloroauric acid, mixing uniformly, adding 25 mu L of chloroplatinic acid solution, mixing uniformly and stirring for 2 minutes; and secondly, adding 27 mu L of 1M sodium hydroxide solution into the solution, uniformly mixing, adjusting the pH value of the solution to 13, and heating in a water bath at 55 ℃ for 10 hours to obtain the chicken protein-gold platinum nanocluster.
Example three: weighing 5mg of chicken protein, and adding 100 mu L of water for dissolving; adding 100 mu L of chloroauric acid, mixing uniformly, adding 12.5 mu L of chloroplatinic acid solution, mixing uniformly and stirring for 2 minutes; and secondly, adding 27 mu L of 1M sodium hydroxide solution into the solution, uniformly mixing, adjusting the pH value of the solution to 13, and heating in a water bath at 55 ℃ for 10 hours to obtain the chicken protein-gold platinum nanocluster.
Example four: the chicken protein-gold platinum nanoclusters obtained in the first embodiment, the second embodiment and the third embodiment are determined to have the strongest fluorescence emission intensity when the concentration ratio of chloroauric acid to chloroplatinic acid is 6:1 by comparing the fluorescence intensity of the chicken protein-gold platinum nanoclusters at an excitation wavelength of 360nm as shown in fig. 2.
Example five: weighing 5mg of chicken protein, and adding 100 mu L of water for dissolving; adding 100 mu L of chloroauric acid, mixing uniformly, adding 16.7 mu L of chloroplatinic acid solution, mixing uniformly and stirring for 2 minutes; and secondly, adding 13 mu L of 1M sodium hydroxide solution into the solution, uniformly mixing, adjusting the pH value of the solution to 9, and heating in a water bath at 55 ℃ for 10 hours to obtain the chicken protein-gold platinum nanocluster.
Example six: weighing 5mg of chicken protein, and adding 100 mu L of water for dissolving; adding 100 mu L of chloroauric acid, mixing uniformly, adding 16.7 mu L of chloroplatinic acid solution, mixing uniformly and stirring for 2 minutes; and secondly, adding 20 mu L of 1M sodium hydroxide solution into the solution, uniformly mixing, adjusting the pH value of the solution to 11, and heating in a water bath at 55 ℃ for 10 hours to obtain the chicken protein-gold platinum nanocluster.
Example seven: the chicken protein-gold platinum nanoclusters obtained in the first embodiment, the fifth embodiment and the sixth embodiment are determined to have the strongest fluorescence emission intensity when the pH of the solution is 13 by comparing the fluorescence intensity of the chicken protein-gold platinum nanoclusters at an excitation wavelength of 360nm as shown in FIG. 3.
Example eight: weighing 5mg of chicken protein, and adding 100 mu L of water for dissolving; adding 100 mu L of chloroauric acid, mixing uniformly, adding 16.7 mu L of chloroplatinic acid solution, mixing uniformly and stirring for 2 minutes; and secondly, adding 27 mu L of 1M sodium hydroxide solution into the solution, uniformly mixing, adjusting the pH value of the solution to 13, and heating in a water bath at 55 ℃ for 3 hours to obtain the chicken protein-gold platinum nanocluster.
Example nine: weighing 5mg of chicken protein, and adding 100 mu L of water for dissolving; adding 100 mu L of chloroauric acid, mixing uniformly, adding 16.7 mu L of chloroplatinic acid solution, mixing uniformly and stirring for 2 minutes; and secondly, adding 20 mu L of 1M sodium hydroxide solution into the solution, uniformly mixing, adjusting the pH value of the solution to 11, and heating in a water bath at 55 ℃ for 6 hours to obtain the chicken protein-gold platinum nanocluster.
Example ten: example one, example eight, and example nine the resulting chicken protein-gold platinum nanoclusters were compared with their respective fluorescence emission intensities at an excitation wavelength of 360nm as shown in fig. 4, and it was determined that the fluorescence emission intensity was the strongest when the reaction time was 10 hours.
Example eleven: weighing 4mg of chicken protein, and adding 100 mu L of water for dissolving; adding 100 mu L of chloroauric acid, mixing uniformly, adding 16.7 mu L of chloroplatinic acid solution, mixing uniformly and stirring for 2 minutes; and secondly, adding 27 mu L of 1M sodium hydroxide solution into the solution, uniformly mixing, adjusting the pH value of the solution to 13, and heating in a water bath at 55 ℃ for 10 hours to obtain the chicken protein-gold platinum nanocluster.
Example twelve: weighing 3mg of chicken protein, and adding 100 mu L of water for dissolving; adding 100 mu L of chloroauric acid, mixing uniformly, adding 16.7 mu L of chloroplatinic acid solution, mixing uniformly and stirring for 2 minutes; and secondly, adding 27 mu L of 1M sodium hydroxide solution into the solution, uniformly mixing, adjusting the pH value of the solution to 13, and heating in a water bath at 55 ℃ for 10 hours to obtain the chicken protein-gold platinum nanocluster.
Example thirteen: first embodiment, the first embodimentEleven, chicken protein-gold nanoclusters obtained in example twelve, as shown in fig. 5, when the concentration of chicken protein is 50mg ml, it is determined by comparing their respective fluorescence emission intensities at an excitation wavelength of 360nm-1The intensity of fluorescence emission is strongest.
Example fourteen: weighing 5mg of chicken protein, and adding 100 mu L of water for dissolving; adding 100 mu L of chloroauric acid, mixing uniformly, adding 16.7 mu L of chloroplatinic acid solution, mixing uniformly and stirring for 2 minutes; and secondly, adding 27 mu L of 1M sodium hydroxide solution into the solution, uniformly mixing, adjusting the pH value of the solution to 13, and heating in a water bath at 25 ℃ for 10 hours to obtain the chicken protein-gold platinum nanocluster.
Fifteen examples 5mg of egg white was weighed and dissolved in 100. mu.L of water; adding 100 mu L of chloroauric acid, mixing uniformly, adding 16.7 mu L of chloroplatinic acid solution, mixing uniformly and stirring for 2 minutes; and secondly, adding 27 mu L of 1M sodium hydroxide solution into the solution, uniformly mixing, adjusting the pH value of the solution to 13, and heating in a water bath at 37 ℃ for 10 hours to obtain the chicken protein-gold platinum nanocluster.
Example sixteen: in the chicken protein-gold platinum nanoclusters obtained in the first embodiment, the fourteenth embodiment and the fifteenth embodiment, as shown in fig. 6, the fluorescence emission intensity is determined to be the strongest when the reaction temperature is 55 ℃ by comparing the fluorescence emission intensities of the chicken protein-gold platinum nanoclusters at an excitation wavelength of 360 nm.
(II) detection of methotrexate by using chicken protein-gold platinum nanocluster as fluorescent probe
Example seventeen: diluting the chicken protein-gold platinum nanocluster stock solution by using a phosphate buffer solution, adding a methotrexate solution with the final concentration of 100-1000 mu M, incubating for 10 minutes at room temperature, and detecting the fluorescence spectrum under the excitation wavelength of 360nm by using a fluorescence spectrometer. As shown in fig. 7 and 8, the fluorescence intensity was linear with the methotrexate concentration at an excitation wavelength of 360 nm.
As can be seen from the above, at 360nm (E)x1) Under the excitation wavelength, the gold platinum nanocluster shows red fluorescence at 455nm (E)m1) And 640nm (E)m2) Has a fluorescence emission peak; the gold platinum nanocluster serving as an effective environment-friendly fluorescent probe can be used for detecting Methotrexate (MTX) in a solution.
The description and practice of the disclosure herein will be readily apparent to those skilled in the art from consideration of the specification and understanding, and may be modified and modified without departing from the principles of the disclosure. Therefore, modifications or improvements made without departing from the spirit of the invention should also be considered as the protection scope of the invention.
Claims (5)
1. A preparation method of a chicken protein protection gold platinum nano-cluster is characterized by comprising the following specific steps:
s1, weighing a certain amount of egg white, adding 100 mu L of water for dissolving, adding chloroauric acid solution for mixing uniformly, adding chloroplatinic acid solution, and stirring and mixing uniformly for 2 minutes to obtain solution A;
s2, adding a 1M sodium hydroxide solution into the solution A obtained in the step S1, adjusting the pH of the solution to 9-13, and heating the mixed solution in a water bath for 3-10 hours to obtain a solution B;
s3, storing the solution B obtained in the step 2 at 4 ℃ to obtain the chicken protein-gold platinum nanocluster.
2. The method for preparing the chicken protein protected gold and platinum nanoclusters of claim 1, wherein in S1, the concentration ratio of chloroauric acid to chloroplatinic acid is 6:1, and the chicken protein concentration is 50mg ml-1。
3. The method of claim 1, wherein in S2, pH of the solution is adjusted to 13 by sodium hydroxide, the heating temperature is 55 ℃, and the reaction time is 10 hours.
4. The application of the chicken protein-gold platinum nanoclusters obtained by the preparation method of chicken protein protection gold platinum nanoclusters as claimed in any one of claims 1 to 3 in methotrexate detection is characterized in that the chicken protein-gold platinum nanoclusters are diluted by phosphate buffer, methotrexate with different concentrations is added and uniformly mixed, the mixture is incubated at room temperature, and under the condition of excitation wavelength, the fluorescence intensity of the chicken protein-gold platinum nanoclusters is gradually reduced along with the gradual increase of the concentration of the methotrexate, so that the detection is realized.
5. The use of the chicken protein-protected gold platinum nanoclusters in methotrexate detection according to claim 4, wherein the detection is realized under the excitation wavelength of 360nm after 10 minutes of incubation at room temperature.
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