CN110976906B - Fluorescent palladium nanocluster and synthesis method and application thereof - Google Patents
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Abstract
The invention provides a fluorescent palladium nanocluster and a synthesis method and application thereof. The palladium nanocluster is prepared by the following method: the fluorescent palladium nanocluster is prepared by one step through a chemical reduction method by taking palladium nitrate as a raw material, polyethyleneimine as a stabilizer and ascorbic acid as a reducing agent. The palladium nanocluster has excellent fluorescence emission performance in an aqueous solution and good photobleaching resistance and stability, and the solution emits bright blue fluorescence under the irradiation of a 365nm ultraviolet lamp. The invention also provides a method for detecting terramycin by using the palladium nanocluster, the terramycin is detected by a fluorescence analysis method, the fluorescence intensity of the palladium nanocluster is gradually reduced along with the increase of the concentration of the terramycin, and meanwhile, the method has good selectivity. The detection method provided by the invention is simple to operate, high in detection speed, good in selectivity and extremely low in cost, and can greatly shorten the detection time.
Description
Technical Field
The invention relates to a metal nano-cluster, and particularly belongs to a fluorescent palladium nano-cluster and a synthetic method and application thereof.
Background
The fluorescent nanoclusters are composed of several to hundreds of atoms and have many characteristics which are not possessed by traditional organic fluorescent molecules, including low toxicity, biocompatibility, large Stokes shift, photobleaching resistance and the like. Besides showing many optical properties similar to molecules, the fluorescent nanoclusters are also important characteristics of large specific surface area, easy surface modification, adjustable emission wavelength and the like. The fluorescent nanocluster has wide application in the aspects of biological detection, biosensors, catalysis, biological imaging and the like, and therefore, the fluorescent nanocluster has a wide application prospect. Currently, fluorescent nanoclusters can be classified into gold (Au), silver (Ag), copper (Cu), palladium (Pd), platinum (Pt), and the like, according to the constituent atoms. In recent years, palladium nanoclusters have mature application in the field of catalysis of organic synthesis, but no report is made on fluorescent palladium nanoclusters as antibiotic sensors.
The existing synthesis method of the fluorescent nanocluster comprises the following steps:
1. the physical method is a method for preparing the nano-cluster by irradiating the metal ion and ligand mixed solution by using ultraviolet light, and the prepared nano-cluster has lower stability and low fluorescence intensity.
2. The chemical reduction method is characterized in that metal ions are reduced under the protection of a stabilizing agent to synthesize metal nano-clusters, the reaction conditions are mild, and the synthesized nano-clusters are good in stability, good in biocompatibility, uniform in size and high in fluorescence intensity.
3. The synthesis method, the microwave method and the ultrasonic method are used for synthesizing the nano-cluster, although the reaction rate is high, the uncontrollable and controllable reaction conditions cause the size of the synthesized nano-cluster to be uneven and the stability to be poor.
Disclosure of Invention
The invention aims to provide a fluorescent palladium nanocluster as well as a synthesis method and application thereof. The application is to use the fluorescent palladium nanocluster for detecting the oxytetracycline, has the performances of good selectivity, high sensitivity, good reproducibility and extremely low cost, and can realize the rapid detection of the oxytetracycline in a tap water sample.
The invention provides a synthesis method of a fluorescent palladium nanocluster, which comprises the following steps:
(1) weighing palladium nitrate, polyethyleneimine and ascorbic acid according to the mass ratio of 1:0.1-30:1-7, ultrasonically dissolving in deionized water, transferring to a round-bottom flask, placing the round-bottom flask in a magnetic stirrer, heating at 40-100 ℃ for 1-12h, and naturally cooling to room temperature after the reaction is finished; filtering the obtained solution with a microporous membrane with the aperture of 0.22 μm, dialyzing with a 1000Da dialysis bag for 48h, removing impurities to obtain yellow palladium nanocluster solution, and storing at 4 deg.C for later use.
The mass ratio of the palladium nitrate, the polyethyleneimine and the ascorbic acid is preferably 1:9: 4.
The heating temperature is preferably 60 ℃, and the heating time is preferably 6 h.
The synthesized fluorescent palladium nanocluster can be used for detecting oxytetracycline.
The invention provides a method for detecting terramycin, which comprises the following specific steps:
(1) preparing a series of terramycin solutions with different concentrations;
(2) setting an excitation wavelength of 360nm and slits of 10nm on an F-4500 fluorescence spectrophotometer, and taking the synthesized palladium nanocluster solution to test the fluorescence intensity of the solution, wherein the emission wavelength of the solution is 495 nm;
(3) adding oxytetracycline solutions with different concentrations into the synthesized palladium nanocluster solution to test the fluorescence intensity of the palladium nanocluster solution, wherein the fluorescence intensity of the palladium nanocluster solution is reduced along with the increase of the concentration of the oxytetracycline solution, and the ratio of the fluorescence intensity is I/I0Establishing a standard curve with terramycin solutions with different concentrations (I is the fluorescence intensity of the palladium nanocluster solution measured by adding the terramycin solution, I0Is the fluorescence intensity of the palladium nanocluster solution measured without adding the oxytetracycline solution).
(4) The fluorescence intensity of the oxytetracycline added in the tap water sample is tested by the palladium nanocluster solution, the concentration of the oxytetracycline added in the tap water sample is calculated according to the established linear relation of the standard curve, and then the recovery rate of the oxytetracycline is calculated.
Compared with the prior art, the invention has the beneficial effects that:
the palladium nanocluster synthesized by the method disclosed by the invention has high stability, can stably exist under various extreme pH values and high ionic strength, has excellent light resistance under ultraviolet irradiation, and is good in biocompatibility, uniform in size and high in fluorescence intensity.
The method for detecting terramycin provided by the invention is based on the principle that palladium nanoclusters are used as energy donors, terramycin is used as energy acceptors, and fluorescence quenching of fluorescence resonance energy transfer occurs between the palladium nanoclusters and the terramycin, so that the rapid detection of the terramycin is realized.
The detection method has the advantages of good selectivity, high sensitivity, simple operation and high detection speed, can greatly shorten the detection time, and can realize the rapid detection of the oxytetracycline in the tap water sample.
Drawings
FIG. 1 is an emission spectrum (A) of a palladium nanocluster solution at different excitation wavelengths and a fluorescence spectrum (B) under 365nm ultraviolet irradiation
FIG. 2 shows selection experiment (A) and detection (B) of oxytetracycline
Detailed Description
Example 1 preparation of palladium nanoclusters:
the specific operation method for synthesizing the palladium nanocluster comprises the following steps: 0.1mL of palladium nitrate 100mM) solution was added to 2.1mL of polyethyleneimine (100mM) solution at 60 ℃ and stirred at a constant speed for 10min, and then 100. mu.L of ascorbic acid (1.5M) was added and reacted for 6 hours. Filtering the obtained solution with microporous membrane with pore diameter of 0.22 μm, centrifuging at 8000rpm/min for 15min, dialyzing with 1000Da dialysis bag for 48 hr, and removing impurities. The resulting solution was stored at 4 ℃ until use.
On an F-4500 fluorescence spectrophotometer, setting an excitation wavelength from 320nm to 400nm, and measuring emission spectra under different excitation wavelengths (shown in figure 1A) by setting slits to be 10nm, wherein 1 to 9 are fluorescence spectra under excitation of 320nm, 330nm, 340nm, 350nm, 360nm, 370nm, 380nm, 390nm and 400nm, and the palladium nanocluster solution emits bright blue fluorescence under 365nm ultraviolet lamp irradiation (shown in figure 1B).
Example 2 Selectivity experiments on oxytetracycline
(1) Ethanol solutions containing ciprofloxacin (Cip), tetracycline (Tc), chloramphenicol (Chl) and erythromycin (Em) were prepared at 1X 10 times respectively-4M; aqueous solutions of amoxicillin (Amx) and kanamycin sulfate (Km) were each 1X 10-4M; preparing an ethanol solution containing oxytetracycline (Ot);
(2) taking the palladium nanocluster solution prepared in the embodiment 1, respectively dropwise adding ciprofloxacin, tetracycline, chloramphenicol, erythromycin, amoxicillin, kanamycin sulfate solution and oxytetracycline ethanol solution, and testing;
(3) on an F-4500 fluorescence spectrophotometer, setting an excitation wavelength of 360nm, and setting a slit to be 10nm, and testing the fluorescence intensity of a palladium nanocluster at 495nm, the fluorescence of the palladium nanocluster is not quenched due to the existence of tetracycline, ciprofloxacin, erythromycin, chloramphenicol, amoxicillin and kanamycin sulfate solution, and only the fluorescence is quenched when oxytetracycline solution is added (as shown in FIG. 2A). Therefore, the palladium nanocluster has good selectivity on oxytetracycline.
Example 3 establishment of a Standard Curve for the detection of oxytetracycline
(1) Preparing a series of terramycin solutions with different concentrations;
(2) on an F-4500 fluorescence spectrophotometer, setting an excitation wavelength of 360nm, wherein slits are all 10nm, and taking the palladium nanocluster solution as in claim 1 to test the fluorescence intensity, wherein the emission wavelength is 495 nm;
(3) adding oxytetracycline solutions with different concentrations into the palladium nanocluster solution to test the fluorescence intensity of the oxytetracycline solutions, wherein the fluorescence intensity of the palladium nanocluster solution is reduced along with the increase of the concentration of the oxytetracycline solution, and the ratio of the fluorescence intensity is I/I0A standard curve (as shown in FIG. 2B) was established for different concentrations of oxytetracycline solution (I is the fluorescence intensity of the palladium nanocluster solution measured by adding oxytetracycline solution, I0Is the fluorescence intensity of the palladium nanocluster solution measured without adding the oxytetracycline solution).
Example 4 recovery Experimental assay for oxytetracycline
(1) Filtering a water sample by using a filter membrane of 0.22 mu m, and adding the water sample into a palladium nano-cluster solution to measure the fluorescence intensity of the water sample;
(2) adding oxytetracycline into the tap water sample to prepare a solution;
(3) the oxytetracycline solution was added to the palladium nanocluster solution, the fluorescence intensity was measured, the oxytetracycline concentration was calculated from the linear relationship of the standard curve, and the recovery rate was calculated (see table 1).
TABLE 1 detection of oxytetracycline in tap water samples
Claims (3)
1. The application of a fluorescent palladium nano-cluster solution in terramycin detection; the fluorescent palladium nanocluster is prepared by a synthesis method comprising the following steps: weighing palladium nitrate, polyethyleneimine and ascorbic acid according to the mass ratio of 1:0.1-30:1-7, ultrasonically dissolving in deionized water, transferring to a round-bottom flask, placing the round-bottom flask in a magnetic stirrer, heating at 40-100 ℃ for 1-12h, and naturally cooling to room temperature after the reaction is finished; filtering the obtained solution with a microporous membrane with the aperture of 0.22 μm, dialyzing with a 1000Da dialysis bag for 48h, removing impurities to obtain yellow palladium nanocluster solution, and storing at 4 deg.C for later use.
2. The use of the fluorescent palladium nanocluster solution as claimed in claim 1 in oxytetracycline detection, wherein the mass ratio of palladium nitrate, polyethyleneimine and ascorbic acid is 1:9: 4.
3. The application of the fluorescent palladium nanocluster solution in the oxytetracycline detection as claimed in claim 1, wherein the heating temperature is 60 ℃ and the heating time is 6 hours.
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CN108778497A (en) * | 2016-03-11 | 2018-11-09 | 东洋合成工业株式会社 | Load type metal catalyst |
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