CN112098382B - Ratio fluorescent probe and application thereof in penicillamine detection - Google Patents
Ratio fluorescent probe and application thereof in penicillamine detection Download PDFInfo
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- CN112098382B CN112098382B CN202010991714.9A CN202010991714A CN112098382B CN 112098382 B CN112098382 B CN 112098382B CN 202010991714 A CN202010991714 A CN 202010991714A CN 112098382 B CN112098382 B CN 112098382B
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- 229960001639 penicillamine Drugs 0.000 title claims abstract description 54
- LJRDOKAZOAKLDU-UDXJMMFXSA-N (2s,3s,4r,5r,6r)-5-amino-2-(aminomethyl)-6-[(2r,3s,4r,5s)-5-[(1r,2r,3s,5r,6s)-3,5-diamino-2-[(2s,3r,4r,5s,6r)-3-amino-4,5-dihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-6-hydroxycyclohexyl]oxy-4-hydroxy-2-(hydroxymethyl)oxolan-3-yl]oxyoxane-3,4-diol;sulfuric ac Chemical compound OS(O)(=O)=O.N[C@@H]1[C@@H](O)[C@H](O)[C@H](CN)O[C@@H]1O[C@H]1[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](N)C[C@@H](N)[C@@H]2O)O[C@@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)N)O[C@@H]1CO LJRDOKAZOAKLDU-UDXJMMFXSA-N 0.000 title claims abstract description 51
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- G—PHYSICS
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- 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"
- G01N21/643—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" non-biological material
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/08—Luminescent, e.g. electroluminescent, chemiluminescent materials containing inorganic luminescent materials
- C09K11/58—Luminescent, e.g. electroluminescent, chemiluminescent materials containing inorganic luminescent materials containing copper, silver or gold
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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
- G01N21/6402—Atomic fluorescence; Laser induced fluorescence
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
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- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/14—Macromolecular compounds
- C09K2211/1408—Carbocyclic compounds
- C09K2211/1425—Non-condensed systems
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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"
- G01N2021/6432—Quenching
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- Chemical Kinetics & Catalysis (AREA)
- Inorganic Chemistry (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
- Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
The invention discloses a ratio fluorescent probe and application thereof in penicillamine detection. The ratiometric fluorescence sensor of the present invention has two emission wavelengths, and the target content is quantified according to the relationship between the ratio of fluorescence intensity at the two wavelengths and the analyte concentration. By establishing an internal standard, the method has a self-regulating function, and greatly weakens the interference of variable or difficult-to-quantify factors on experiments and results. Moreover, as the concentration of the analyte changes, the color of the ratio probe also changes stepwise, thereby realizing the visual detection of the analyte, and semi-quantitative and qualitative analysis of the analyte can be realized according to the color of the probe.
Description
Technical Field
The invention relates to a ratio fluorescent nano-probe phenolic resin-Au cluster-Cu 2+ detection system and construction thereof, and also relates to application of the ratio fluorescent nano-probe in detecting penicillamine.
Background
Penicillamine (D-PENICILLAMINE, D-PA) chemical name 3, 3-dimethyl- (D) -cysteine is an amino acid containing sulfhydryl group, and has the capability of chelating metal ions, wherein the effect of chelating Cu 2+ is particularly remarkable. Therefore, it has outstanding capability in treating hepatolenticular degeneration (HLD, a copper metabolic disorder), heavy metal poisoning and the like. On the other hand, the mercapto group of D-penicillamine can decompose rheumatoid factors belonging to macroglobulin due to disulfide bond cleavage, thereby reducing the level of serum rheumatoid factors. Based on this, penicillamine has been widely used for autoimmune diseases such as scleroderma and rheumatoid arthritis. In addition, penicillamine can interfere with cross-linking to collagen (Tropocollagen) to form insoluble collagen tissue and prevent the maturation of soluble collagen, so it can be used to promote connective tissue proliferation. Penicillamine also has an anti-inflammatory effect, mainly manifested by inhibition of the release of lysosomal enzymes, to stabilize lysosomal membranes. In addition Xu Hong et al found that by studying mice, high doses of penicillamine, while achieving the desired effect of reducing copper levels in vivo, also caused some side effects; the effect of the low dose of penicillamine is not great; while moderate doses of penicillamine can not only significantly remove copper elements in serum and liver, but also have no adverse effect on experimental rats. Therefore, the method for simply and rapidly detecting the concentration of the penicillamine in the medicine and the body has great significance for the research on the aspects of biomedicine and pharmacology.
Fluorescent metal nanoclusters composed of several metal atoms are a promising fluorescent probe due to their small size, good light stability, customizable surface properties and high photoluminescence depending on the size, and a wide range of applications. Compared with silver and copper nanoclusters which are easy to oxidize, the luminescent gold nanoclusters are stable in chemical property and easy to control in size. Gold nanoclusters have therefore recently been considered as a satisfactory candidate for imaging, detecting metal ions, and for manufacturing fluorescence sensors.
At present, various methods for determining the content of penicillamine exist. Such as high performance liquid chromatography, infrared spectroscopy, ultraviolet spectrophotometry, and the like. However, the above method, such as high performance liquid chromatography, has the disadvantages of high cost, small detection amount, need of using different types of filling columns, difficulty in analyzing inorganic ions and biological macromolecules, large consumption of mobile phase, toxicity, and out-of-column effect. The infrared spectrometry has the defects that the water-containing sample is unsuitable to be analyzed because the hydroxyl peak in water has larger interference on measurement, the error is larger when quantitative analysis is performed, the sensitivity is lower, and experience is mainly relied on when the analysis of the atlas is performed. The ultraviolet spectrophotometry is expensive in equipment, ultraviolet absorption of different substances is different, certain errors exist in measurement results, and experimental results are influenced and limited by factors such as cuvette materials, pH values of solutions, buffer medium solutions, light sources and the like. Although in recent years, with the continuous progress of fluorescein and fluorescence detection technology, the sensitivity of fluorescence detection has become very high, the level of isotope labeling detection is reached, and compared with the isotope labeling method, the fluorescence labeling method has the advantages of safety, convenience, no radioactive pollution and the like, so that the concentration of penicillamine can be detected by using a fluorescence probe method. However, penicillamine molecules are themselves non-fluorescent, and thus derivatized fluorescence spectrometry to determine penicillamine is an option. However, the derivatization method is difficult to popularize in practical application due to complicated operation, low sensitivity, poor reproducibility and the like, and based on the reasons, a brand new method is urgently needed to make up for various defects of the method so as to achieve rapid and accurate detection of penicillamine. Single-emission signal fluorescence sensors have considerable advantages to make up for the deficiencies of some conventional approaches. However, the single-emission signal fluorescence sensor has defects, such as that the quantum dot nano probe is easily interfered by a plurality of variable or difficult-to-quantify factors such as probe concentration, polarity, temperature, pH value of environment, stability and the like, so that the uncertainty of the signal is caused.
Disclosure of Invention
The invention provides a ratio fluorescent probe phenolic resin-Au cluster-Cu 2+ detection system and application thereof in penicillamine detection, which aims to solve the defects of the existing penicillamine detection technology.
According to the invention, the ratio fluorescence sensor phenolic resin-Au quantum dot with double fluorescence emission is researched and constructed, the sensor has a quenching effect on the fluorescence of the Au quantum dot through Cu 2+, and penicillamine can interact with Cu 2+, so that the quenching of Cu 2 + is relieved, and the Au quantum dot gradually recovers the fluorescence, so that the detection of D-penicillamine with high sensitivity and high accuracy is realized. Whereas the fluorescent phenolic resin (PFR) as an internal standard is not responsive to Cu 2+ and penicillamine. Therefore, the PFR is taken as an internal standard, and the fluorescence intensity of the Au quantum dot is observed, so that the linear relation between the ratiometric fluorescence and the penicillamine concentration is made.
The ratiometric fluorescence sensor of the present invention has two emission wavelengths, and the target content is quantified according to the relationship between the ratio of fluorescence intensity at the two wavelengths and the analyte concentration. By establishing an internal standard, the method has a self-regulating function, and greatly weakens the interference of variable or difficult-to-quantify factors on experimental results. Moreover, as the concentration of the analyte changes, the color of the ratio probe also changes step by step, thereby realizing the visual detection of the analyte, improving the detection legibility and the detection speed, and realizing the semi-quantitative and qualitative analysis of the analyte according to the color of the probe. The established sensor mode improves the range of dynamic response by means of the change of the fluorescence intensity ratio.
If only a single fluorescent probe of an Au cluster is used for detecting penicillamine, the orange-red fluorescence with single fluorescence intensity change is difficult to distinguish directly by naked eyes (the orange-red fluorescence fades down with the decrease of the penicillamine concentration, and the fading down of single color is not easy to distinguish by naked eyes); the invention adopts the ratio fluorescent probe of phenolic resin-Au cluster, and the blue-green light of the fluorescent phenolic resin gradually changes from red to blue along with the decrease of the concentration of penicillamine, so that the color change which is easy to distinguish appears, as shown in figure 4. Therefore, compared with a single fluorescence intensity change detection method, the ratio fluorescence detection method is more sensitive, and visual detection is more reliable and easy to distinguish.
Drawings
FIG. 1 is a graph showing fluorescence emission curves (610 nm emission peak) of the prepared Au clusters under 270nm excitation light and ultraviolet-visible absorption curves thereof.
FIG. 2 is a graph showing fluorescence emission curves (410 nm emission peaks) and ultraviolet-visible absorption curves of the prepared phenolic resin under 312nm excitation light.
FIG. 3 is a graph showing fluorescence emission curves of Cu 2+ quenched Au clusters in the construction of a system for detecting penicillamine solution.
FIG. 4 is a graph showing the recovery of Au cluster fluorescence by penicillamine when penicillamine was detected in a phenolic resin-Au cluster-Cu 2+ solution system.
FIG. 5 is a graph showing the relationship between penicillamine concentration and the ratio of Au cluster fluorescence intensity (I 610/I410) to phenolic resin.
FIG. 6 shows the selectivity of the phenolic-Au cluster-Cu 2+ detection system for penicillamine.
Fig. 7 is an SEM image of fluorescent phenolic resin.
Detailed Description
Example 1
The preparation method of the Au cluster comprises the following steps:
Firstly, accurately preparing 0.01M chloroauric acid solution (after 1g chloroauric acid is dissolved in deionized water, a 250mL volumetric flask is used for constant volume); 200mL of 1M NaOH solution was then prepared for further use. Into a 50mL round bottom flask was added 10mL of deionized water, 0.0544g of MUA (11-mercaptoundecanoic acid) was weighed and added to the round bottom flask, after 5min of sonication, 1M NaOH solution was added while sonicating to completely dissolve the MUA, and the addition of NaOH was stopped. Subsequently 6250. Mu.L of the previously prepared HAuCl 4 solution, if floc is present, was added slowly with stirring, stirred for half an hour, if the precipitate had not disappeared, naOH solution (1M) was added dropwise until the solution became clear, and stirred at room temperature for 24h. And dialyzing the reacted solution for 24 hours, obtaining a colorless solution which is strongly orange-red light in 312nm ultraviolet light, namely an Au cluster solution, centrifuging at 10000rpm for 10 minutes before use, discarding white precipitate, and taking supernatant for use.
Example 2
The preparation method of the phenolic resin comprises the following steps:
0.05mmol of phenol and 0.025mmol of Hexamethylenetetramine (HMT) are dispersed in 22ml of ultrapure water, and after being fully dissolved, the mixture is transferred into a hydrothermal reaction kettle for reaction for 4 hours at a constant temperature of 160 ℃. And (5) preserving the product obtained by the reaction at low temperature. The obtained phenolic resin aqueous solution shows bluish green light in 312nm ultraviolet light. Before the experiment, the inner container of the reaction kettle is soaked in a mixture of concentrated HNO 3 and H 2 O in a volume ratio of 10:1 for 4 hours, then ultrapure water is added for constant temperature 140 ℃ for cleaning for 4 hours, and finally the inner container is dried. From FIG. 7, the phenolic resin of the present invention is uniformly distributed and uniform in size.
Example 3
The construction method of the solution system for detecting penicillamine by ratio fluorescence is as follows:
① To accurately prepare 10mM penicillamine, 1000. Mu.L of 10mM penicillamine and 9.0ml of deionized water were pipetted to prepare 1.0mM penicillamine for use.
② Accurately prepare 10mM Cu (NO 3)2 solution), aspirate 1000. Mu.L of 10mM Cu (NO 3)2 solution with 9.0mL of deionized water to prepare 1.0mM Cu (NO 3)2 solution) with a 1000. Mu.L pipette.
③ To a test tube was added 2mL of water, 1mL of Au cluster solution, and 1mL of aqueous phenolic resin solution, for use.
④ 2ML of water and 100. Mu.L of a phenolic resin-Au cluster mixed solution are added into a cuvette, 2, 4, 6,8, 10, 12, 14, 16, 18, 20, 22, 24, 26 and 28. Mu.L of a 1mM Cu (NO 3) 2 solution are sequentially added, the concentration of Cu (NO 3) 2 in a detection system is sequentially 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 and 14. Mu.M, fluorescence spectra are sequentially recorded, and the fluorescence intensity of the Au clusters is not reduced after 28. Mu.L of the 1mM Cu (NO 3) 2 solution is added, namely the phenolic resin-Au cluster-Cu 2+ detection system (ratio fluorescent probe) is shown in FIG. 3. Copper ions are added to quench the Au cluster fluorescence in the fluorescent probe.
Example 4
The detection system of phenolic resin-Au cluster-Cu 2+ is used for detecting penicillamine, and the method is as follows:
Detecting by adopting a fluorescence spectrum method, taking a phenolic resin-Au cluster-Cu 2+ solution detection system configured in the embodiment 3, sequentially adding 2, 4, 6, 8, 10, 12, 14, 16, 18 and 20 mu L of 1mM penicillamine, sequentially enabling the concentration of the penicillamine in the detection system to reach 1,2,3, 4, 5, 6, 7, 8, 9 and 10 mu M, and sequentially recording the fluorescence spectrum of the solution system, wherein the fluorescence spectrum is shown in FIG. 4; the relationship between the ratio of the fluorescence intensity of the Au cluster to the fluorescence intensity of the phenol resin and the concentration of penicillamine was obtained from the fluorescence spectrum, as shown in FIG. 5.
Example 5
The selective detection experiment of the phenolic resin-Au cluster-Cu 2+ detection system on penicillamine is as follows;
1mM of L-lysine, L-valine, D-alanine, L-leucine, L-glutamic acid, glucose, naCl, KCl, caCl 2 and penicillamine solutions are respectively prepared, 5 mu L of each solution is respectively and independently added into a phenolic resin-Au cluster-Cu 2+ solution detection system prepared in example 3, a fluorescence spectrum method is adopted, corresponding fluorescence spectrograms are respectively recorded, the change delta I of fluorescence intensity of Au clusters before and after the solution is added is recorded, and the result is that compared with other solutions, the phenolic resin-Au cluster-Cu 2+ detection system (ratio fluorescent probe) has the most sensitive detection response to penicillamine.
Example 6
The detection system of phenolic resin-Au cluster-Cu 2+ is used for detecting the concentration of penicillamine in urine, and the method is as follows:
Taking 2mL of urine sample into a cuvette, adding 100 mu L of phenolic resin-Au cluster mixed solution (prepared in example 3) and 28 mu L of 1mM Cu (NO 3)2 solution, respectively and independently adding 4, 10 and 16 mu L of 1mM penicillamine solution to enable the concentration of penicillamine in the system to reach 2, 5 and 8 mu M, adopting a fluorescence spectrometry method to record corresponding spectrograms respectively, calculating the fluorescence intensity ratio of the Au cluster to the phenolic resin according to the spectrograms, calculating the fluorescence intensity ratio of the Au cluster to the phenolic resin according to the relation between the obtained penicillamine concentration and the fluorescence intensity ratio of the Au cluster to the phenolic resin when the penicillamine concentration reaches 2, 5 and 8 mu M, and comparing the fluorescence intensity ratio corresponding to the penicillamine concentration in urine with the ratio.
TABLE 1 penicillamine detection assay in urine
| Sample | Add(μM) | Found(μM) | Recovery(％) |
| 1 | 2.0 | 2.27 | 113.52 |
| 2 | 5.0 | 5.23 | 104.63 |
| 3 | 8.0 | 8.12 | 101.54 |
It should be noted that the foregoing technical disclosure is only for explanation and illustration to enable one skilled in the art to know the technical spirit of the present invention, and the technical disclosure is not intended to limit the scope of the present invention. The essential scope of the invention is as defined in the appended claims. Those skilled in the art should understand that any modification, equivalent substitution, improvement, etc. made based on the spirit of the present invention should fall within the spirit and scope of the present invention.
Claims (8)
1. The application of the ratio fluorescent probe in penicillamine detection is characterized in that the ratio fluorescent probe is a detection system mainly composed of phenolic resin, au clusters and Cu 2+ in a compounding way; the preparation method of the ratio fluorescent probe comprises the steps of synthesizing fluorescent phenolic resin, preparing Au clusters and constructing a phenolic resin-Au cluster-Cu 2+ detection system;
The synthesis of the fluorescent phenolic resin adopts phenol and hexamethylenetetramine to carry out hydrothermal reaction for 3-6h at 150-180 ℃;
The construction of the phenolic resin-Au cluster-Cu 2+ detection system comprises the following steps: uniformly mixing an Au cluster solution and a phenolic resin solution to obtain an Au cluster-phenolic resin mixed solution for later use; and respectively taking equal amounts of Au cluster-phenolic resin mixed solution, sequentially adding Cu 2+ solutions with the same concentration and different amounts, and sequentially recording fluorescence spectrograms until the fluorescence intensity of the Au cluster is no longer reduced, thus obtaining the phenolic resin-Au cluster-Cu 2+ detection system.
2. The use according to claim 1, wherein the fluorescent phenolic resin is synthesized by hydrothermal reaction of phenol with hexamethylenetetramine at 160 ℃ for 4 hours.
3. The use according to claim 1, wherein the molar ratio of phenol to hexamethylenetetramine is from 1.8 to 2.2:1.
4. The use according to claim 3, wherein the molar ratio of phenol to hexamethylenetetramine is 2:1.
5. The application of claim 1, wherein the inner container of the reaction kettle for the hydrothermal reaction is soaked in a mixture of concentrated HNO 3 and H 2 O in a volume ratio of 10:1 for 4-5 hours, then ultrapure water is added to clean the inner container for 4-5 hours at a constant temperature of 140-150 ℃, and the inner container is dried and then subjected to the hydrothermal reaction.
6. The use of claim 1, wherein the Au-clusters are prepared by reacting 11-mercaptoundecanoic acid with chloroauric acid solution.
7. The use of claim 1, wherein the method of preparing the ratiometric fluorescent probe comprises:
(1) Synthesis of fluorescent phenolic resin: 0.05 Dispersing mmol phenol and 0.025 mmol hexamethylenetetramine in 22 ml ultrapure water, fully dissolving, transferring into a hydrothermal reaction kettle, reacting at 160 ℃ for 4 hours, and preserving the obtained product at low temperature;
(2) Preparation of Au clusters: preparing 0.01M chloroauric acid solution, dissolving 1g chloroauric acid in deionized water, and fixing the volume in a 250mL volumetric flask; preparing 1M NaOH solution 200 mL for later use; adding 10mL of deionized water into a 50mL round-bottom flask, weighing 0.0544-0.0546 g of 11-mercaptoundecanoic acid, adding into the round-bottom flask, after ultrasonic treatment for 5: 5 min, adding 1: 1M NaOH solution while ultrasonic treatment to completely dissolve MUA, stopping adding NaOH, slowly adding 6250 mu L of the previously prepared HAuCl 4 solution while stirring, if floccules appear, stirring for half an hour, precipitating if floccules do not disappear, dripping 1M NaOH solution until the solution becomes clear, stirring at room temperature for 24: 24h, dialyzing the reacted solution for 24: 24h, wherein the obtained solution is colorless under sunlight, and is strongly orange-red and shiny under 312: 312 nm ultraviolet light, namely the Au cluster;
(3) Construction of a phenolic resin-Au cluster-Cu 2+ detection system:
Taking the Au cluster solution in the step (2) and the phenolic resin solution in the step (1), and uniformly mixing to obtain an Au cluster-phenolic resin mixed solution for later use; and respectively taking equal amounts of Au cluster-phenolic resin mixed solution, sequentially adding 1.0 mM of Cu 2+ solutions with different amounts, and sequentially recording fluorescence spectrograms until the fluorescence intensity of the Au cluster is no longer reduced, thus obtaining the phenolic resin-Au cluster-Cu 2+ detection system.
8. The use according to claim 1, wherein the ratio of the fluorescence intensity of the Au clusters to the fluorescence intensity of the phenolic resin is obtained by taking an equal amount of the phenolic resin-Au cluster-Cu 2+ solution detection system by fluorescence spectrometry, sequentially adding 1 mM penicillamine in different amounts, sequentially recording the fluorescence spectrum of the solution system, and obtaining the relationship between the concentration of penicillamine and the ratio of the fluorescence intensity of the Au clusters to the fluorescence intensity of the phenolic resin according to the fluorescence spectrum.
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| A fluorescence detection of D-penicillamine based on Cu2+-induced fluorescence quenching system of protein-stabilized gold nanoclusters;Peng Wang et.al.;《Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy》;参见第199页左栏第3段-201页右栏第3段 * |
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