CN109336773A - A kind of preparation method and application of fluorescent optical sensor - Google Patents
A kind of preparation method and application of fluorescent optical sensor Download PDFInfo
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Abstract
The invention discloses a kind of preparation method and application of fluorescent optical sensor, this method synthesizes column [5] aromatic hydrocarbons first, is then used to restore gold chloride and synthesizes gold nanoparticle rPA5-AuNPs as stabilizer；Using column [5] aromatic hydrocarbons as supermolecule main block, rhodamine B is fluorescence indicator, RhB fluorescence is quenched using the golden nanometer particle dispersion liquid of column [5] aromatic hydrocarbons preparation, then RhB is extruded from the cavity of supermolecule by the way that cholesterol is added, so that fluorescence intensity be made to restore；Using RhB@rPA5-AuNPs as fluorescence sense platform, come quantitative detection cholesterol in the way of fluorescence " On-Off-On "；Excessively complicated, detection speed that the present invention overcomes the cholesterol detection methods of the prior art is slowly and to the lower defect of cholesterol identity；The method of the present invention is simple, easily and fast, controllability it is high, efficiently, suitable for industrialized production and have a vast market application prospect.
Description
Technical field
The invention belongs to fluorescent optical sensor fields, and in particular to a kind of preparation side of the fluorescent optical sensor of cholesterol detection
Method and application.
Background technique
Cholesterol (3 β-cholesteric -5- alkene -3- alcohol) is the wax found in the mankind and higher mammal fat, blood and cell
Matter steroid metabolism substance.It is not only a component part of cell membrane, and is bile acid, and vitamin D and steroids swash
The precursor of element synthesis.As a part of healthy diet intake fat, cholesterol plays important work in brain, nerve and in being immunized
With being the important biomolecule mark of several diseases.Cholesterol levels in human serum have become clinical diagnosis and prevention is cardiovascular
The important indicator of disease.Healthy Human Serum cholesterol levels are lower than 5.17 mmol/L, research shows that high-level cholesterol can draw
Play coronary heart disease and peripheral arterial disease, diabetes, hypertension, sudden cardiac arrest and anaemia etc.；And low-level cholesterol may also
Cause some diseases, such as hypolipoproteinemia, obesity, septicemia and malnutrition.Therefore, Cholesterol measurement, which has become, faces
One of the test most often carried out in bed laboratory.
Current most of cholesterol tests use the biosensor based on enzyme.However, the test based on enzyme is expensive
And the catalytic activity for needing to safeguard enzyme, there are also other methods cholesterol detections, such as high performance liquid chromatography, colorimetric method, electricity
Chemical method and electrochemical luminescence etc..But these existing detection methods, there are detecting step complexity, detection speed is slow and solid to gallbladder
The higher problem of alcohol detection limit.
Summary of the invention
Present invention seek to address that existing cholesterol detection method is excessively complicated, detection speed is slow and to cholesterol identity compared with
Low problem, and a kind of preparation method of the fluorescent optical sensor of cholesterol detection is provided.
The present invention synthesizes column [5] aromatic hydrocarbons first, is then used to restore gold chloride and synthesizes gold nanoparticle as stabilizer,
Fluorescence sense RhB@rPA5-AuNPs is constructed, come quantitative detection cholesterol in the way of fluorescence " on-off-on ".It utilizes
Based on rPA5, fluorescence indicator RhB quenches RhB fluorescence using rPA5-AuNPs solution, then solid by the way that gallbladder is added
Alcohol restores RhB fluorescence intensity, and the linear relationship of concentration is added come quantitative detection gallbladder according to fluorescence intensity change and cholesterol
Sterol.
To achieve the above object, the invention is realized by the following technical scheme:
Fluorescent optical sensor of the present invention the preparation method is as follows:
(1) quinhydrones two (2- hydroxyethyl) ether, triphenylphosphine, anhydrous acetonitrile are sequentially placed into flask, it is cooling with ice-water bath, it stirs
After mixing mixing, it is slowly added to carbon tetrabromide, is stirred to react at room temperature, cold water is added into mixture after complete reaction and is quenched instead
It answers, obtains white precipitate, sediment is collected by filtration, washed 3-4 times with methanol aqueous solution, with recrystallizing methanol, is made after dry
Compound 1, wherein the mass ratio of (2- hydroxyethyl) ether of quinhydrones two and triphenylphosphine is 0.1-1:1, (the 2- hydroxyl second of quinhydrones two
Base) mass ratio of ether and carbon tetrabromide is 0.1-0.5:1；
(2) compound 1, paraformaldehyde, 1,2- dichloroethanes are added in flask, and cooling with ice-water bath, then thereto
Boron trifluoride ether is added, is stirred to react at room temperature, after fully reacting plus water quenching is gone out, and is finally extracted with dichloromethane, organic phase
It is dry with anhydrous sodium sulfate, removing solvent is concentrated under reduced pressure and obtains crude product；Crude product obtains compound 2 by column chromatographic purifying,
Wherein the mass ratio of compound 1 and paraformaldehyde is 5-10:1, and compound 1 and the mass ratio of boron trifluoride ether are 0.5-2:
1；
(3) ethyl alcohol, compound 2,2- (dimethylamino) ethyl alcohol are added in flask, back flow reaction 24-72h, after reaction
It is cooled to room temperature, is concentrated under reduced pressure and removes solvent, residual solids are soluble in water, are filtered to remove insoluble matter, the rotation of gained filter vacuum
Solvent is evaporated off, finally by residue ethanol washing, vacuum drying obtains light tan solid column [5] aromatic hydrocarbons (rPA5), wherein
The mass ratio of compound 2 and 2- (dimethylamino) ethyl alcohol is 0.1-0.5:1；
；
(4) PBS buffer solution, chlorauric acid solution, column [5] arene solution are added in water, is placed in mixed solution after stirring and evenly mixing
0.5-1.5h is reacted at 80-120 DEG C, solution is centrifuged after becoming claret, and precipitating is used deionized water eccentric cleaning 3-4 times, is made
RPA5-AuNPs fluorescent optical sensor, wherein the molar ratio of gold chloride and column [5] aromatic hydrocarbons is 0.01-0.05:1, and water and PBS are buffered
The volume ratio of liquid is 6-8:1, and the volume ratio of PBS buffer solution and chlorauric acid solution is 5-6:1.
The methanol aqueous solution is that 3:2 is mixed to prepare first alcohol and water by volume.
The PBS buffer solution is the liquid of 0.1mol/L, pH=7.0.
The concentration of the chlorauric acid solution is 0.01-0.02 mol/L, and column [5] arene solution concentration is 0.02-0.04
mol/L。
The present invention consolidates another object is that applying fluorescent optical sensor made from the preparation method of above-mentioned fluorescent optical sensor in gallbladder
In alcohol (Cho) detection, come quantitative detection cholesterol in the way of fluorescence " on-off-on ", based on rPA5, fluorescence instruction
Agent is rhodamine B (RhB), RhB fluorescence is quenched using rPA5-AuNPs fluorescent optical sensor solution, then by the way that cholesterol is added
Restore RhB fluorescence intensity, it is solid come quantitative detection gallbladder that the linear relationship of concentration is added according to fluorescence intensity change and cholesterol
Alcohol.
Cholesterol fluorescent optical sensor high sensitivity of the invention；RhB fluorescence is quenched using solution of gold nanoparticles, is then passed through
Cholesterol, which is added, restores RhB fluorescence intensity, and the linear relationship of concentration is added to quantify with cholesterol according to fluorescence intensity change
Cholesterol detection；This composite sensing interface based on RhB@rPA5-AuNPs, than common composite sensing interface, sensitivity is more
Height, stability are more preferable；The method of the present invention carries out at normal temperatures and pressures, is simple, is quick, controllability is high, before wide application
Scape.
Detailed description of the invention
Fig. 1 is compound 11H NMR(A) and13C NMR(B)；
Fig. 2 is compound 21H NMR(A) and13C NMR(B)；
Fig. 3 is compound rPA5's1H NMR(A) and13C NMR(B)；
Fig. 4 is the schematic diagram of the fluorescent optical sensor cholesterol detection constructed based on RhB@rPA5-AuNPs gold nanoparticle；
Fig. 5 is the infrared spectrogram of rPA5 and rPA5-AuNPs；
Fig. 6 is the fluorescence spectra that 4 rPA5-AuNPs of the embodiment of the present invention quenches RhB, and abscissa is wavelength, and ordinate is glimmering
Luminous intensity；
Fig. 7 is that 5 cholesterol of the embodiment of the present invention restores spectrogram to the fluorescence of RhB@rPA5-AuNPs, and abscissa is wavelength, is indulged
Coordinate is fluorescence intensity；
Fig. 8 is the linear relationship schematic diagram of the degree that RhB@rPA5-AuNPs system fluorescence intensity is restored and cholesterol concentration；
Fig. 9 is the binding constant double reciprocal plot of RhB and rPA5, and wherein A figure is fluorescent absorption curve, and B figure is RhB and rPA5
Between binding constant；
Figure 10 is the binding constant double reciprocal plot of Cho and rPA5, and wherein A figure is fluorescent absorption curve, B figure be cholesterol and
Binding constant between rPA5；
Figure 11 is the interference free performance that RhB@rPA5-AuNPs sensor identifies Cho.
Specific embodiment
Further to disclose rather than the present invention is limited, the present invention is made further specifically with reference to embodiments
It is bright.
Chemical reagent used in embodiment and solvent are that analysis is pure；The stirring uses magnetic stirrer side
Formula.The fluorescence spectrometry condition is launch wavelength 530-700nm, excitation wavelength 510nm, and slit width is
10nm。
Embodiment 1: this fluorescent optical sensor the preparation method is as follows:
(1) 10g quinhydrones two (2- hydroxyethyl) ether, 31.5g triphenylphosphine, 250mL anhydrous acetonitrile are sequentially placed into round-bottomed flask
In, it is cooling with ice-water bath, after stirring and evenly mixing, 39.8g carbon tetrabromide is added, is stirred to react 4h at room temperature, after complete reaction to
200mL cold water quenching reaction is added in mixture, obtains white precipitate, sediment is collected by filtration, with methanol aqueous solution (volume ratio
3:2) wash 3 times, with recrystallizing methanol, 14.5 g of white crystal after drying, compound 11H NMR and13C NMR(Fig. 1)；
(2) by 3.37g compound 1,0.349 g paraformaldehyde, 50mL 1,2- dichloroethanes is added in round-bottomed flask, is used in combination
Ice-water bath is cooling, and 3.26g boron trifluoride ether is then added thereto, is stirred to react 1h at room temperature, and 50mL is added after fully reacting
Water quenching is gone out, and is finally extracted with dichloromethane, and organic phase is dry with anhydrous sodium sulfate, and removing solvent is concentrated under reduced pressure and obtains crude product；
Crude product is purified by silica gel column chromatography, and eluent, vacuum refrigeration are collected in petroleum ether-ethyl acetate (volume ratio 100:1) elution
It is dried to obtain compound 2, compound 21H NMR and13C NMR(Fig. 2)；
(3) 30mL ethyl alcohol, 0.2g compound 2,0.85g 2- (dimethylamino) ethyl alcohol are added in round-bottomed flask, back flow reaction
48h is cooled to room temperature after reaction, is concentrated under reduced pressure and is removed solvent, and residual solids are dissolved in 5mL water, is filtered to remove insoluble matter
Matter, gained filter vacuum revolving remove solvent, and finally by residue ethanol washing, vacuum drying obtains light tan solid column
[5] aromatic hydrocarbons 0.22g, rPA5's1H NMR(A) and13C NMR(B) (Fig. 3)；
(4) in 35mL water be added 5mLPBS buffer (0.1 mol/L, pH 7.0), 1mL chlorauric acid solution (0.01mol/L),
Mixed solution, is placed at 100 DEG C after stirring and evenly mixing and reacts 1h, solution becomes wine by 10mL column [5] arene solution (0.02mol/L)
It is centrifuged after red, precipitating is used deionized water eccentric cleaning 3 times, and rPA5-AuNPs fluorescent optical sensor is made.
Embodiment 2: this fluorescent optical sensor the preparation method is as follows:
(1) 10g quinhydrones two (2- hydroxyethyl) ether, 12.5g triphenylphosphine, anhydrous acetonitrile are sequentially placed into flask, use ice water
Bath cooling after stirring and evenly mixing, is added 25g carbon tetrabromide, is stirred to react 5h at room temperature, is added after complete reaction into mixture
Cold water quenching reaction, obtains white precipitate, and sediment is collected by filtration, and is washed 4 times with methanol aqueous solution (volume ratio 3:2), uses first
Compound 1 is made after dry in alcohol recrystallization；
(2) by 3.0g compound 1,0.43g paraformaldehyde, 50mL 1,2- dichloroethanes is added in round-bottomed flask, and uses ice
Then 2.0g boron trifluoride ether is added thereto, is stirred to react 1h at room temperature for water-bath cooling, 50mL water quenching is added after fully reacting
It goes out, is finally extracted with dichloromethane, organic phase is dry with anhydrous sodium sulfate, and removing solvent is concentrated under reduced pressure and obtains crude product；It is thick to produce
Object is purified by silica gel column chromatography, and eluent, vacuum freeze drying are collected in petroleum ether-ethyl acetate (volume ratio 100:1) elution
Obtain compound 2；
(3) 30mL ethyl alcohol, 0.2g compound 2,0.5g 2- (dimethylamino) ethyl alcohol are added in round-bottomed flask, back flow reaction
48h is cooled to room temperature after reaction, is concentrated under reduced pressure and is removed solvent, and residual solids are dissolved in 5mL water, is filtered to remove insoluble matter
Matter, gained filter vacuum revolving remove solvent, and finally by residue ethanol washing, vacuum drying obtains light tan solid column
[5] aromatic hydrocarbons, Fig. 5 infrared spectroscopy explanation, the characteristic peak of rPA5 have 3015.03 cm-1, 2953.69 cm-1, 1613.34 cm-1,
1500.28 cm-1, 1478.91 cm-1, 1403.87 cm-1.Wherein 3015.03 cm-1It is stretched for unsaturation C-H on phenyl ring
Contracting vibration peak； 2953.69 cm-1For the stretching vibration peak for being saturated C-H；1613.34 cm-1, 1500.28 cm-1,
1478.91 cm-1For phenyl ring skeletal vibration absorption peak；1403.87 cm-1For C-H bending vibration absorption peak.
(4) 6mLPBS buffer (0.1 mol/L, pH 7.0), 1mL chlorauric acid solution are added in 36mL water
Mixed solution, is placed at 90 DEG C after stirring and evenly mixing and reacts by (0.01mol/L), 25mL column [5] arene solution (0.02 mol/L)
1.5h, solution are centrifuged after becoming claret, and precipitating is used deionized water eccentric cleaning 4 times, and rPA5-AuNPs fluorescent optical sensor is made
(Fig. 4 is the preparation of rPA5-AuNPs fluorescent optical sensor and uses principle), rPA5-AuNPs infrared spectroscopy illustrates rPA5- in Fig. 5
The characteristic peak of AuNPs has 3454.85 cm-1, 3016.04 cm-1, 2946.21 cm-1, 1739.28 cm-1, 1437.19 cm-1, 1365.84 cm-1。 3454.85 cm-1For the stretching vibration peak of-OH contained by rPA5；3016.04 cm-1For on phenyl ring not
It is saturated the stretching vibration peak of C-H；2946.21 cm-1For the stretching vibration peak for being saturated C-H；1437.19 cm-1,
1365.84 cm-1For C-H bending vibration absorption peak；Compare the difference of two spectrograms, above data illustrate rPA5-AuNPs at
Function preparation.
Embodiment 3: the experiment of the binding constant of rhodamine B and column [5] aromatic hydrocarbons, cholesterol and column [5] aromatic hydrocarbons
The rPA5 solution of 1 μm of ol/L is added by several times into 10 μm of ol/L RhB solution of 2mL, every time add 10 μ l, obtain one group it is glimmering
Absorption curves make corresponding dependent linearity relationship (Fig. 9 A) according to curve, to calculate between RhB and rPA5
Binding constant Ka1(Fig. 9 B).It is molten that 1 μm of ol/L cholesterol is added into 10 μm of ol/L rPA5 solution of 2mL in same method by several times
Liquid adds 10 μ l every time, obtains one group of fluorescent absorption curve (Figure 10 A), make corresponding linear relationship also according to curve, counts
Calculate the binding constant Ka between rPA5 and Cho2(Figure 10 B)；Binding constant Ka between rPA5 and Cho2Greater than RhB with
Binding constant Ka between rPA51；Different competitive relations between the above Subjective and Objective are that Cho squeezes RhB from the cavity of rPA5
Experiment basis has been established out.
RhB fluorescence is quenched in embodiment 4:rPA5-AuNPs fluorescent optical sensor
Using ultrapure water compound concentration is that 800 μm of ol/L RhB are stand-by as stock solution, and in the test tube of 10mL, RhB is added
Stock solution and ultrapure water mix, and the RhB solution with 10 μm of ol/L of obtained 2mL measures its fluorescence intensity；Then into the solution
10 μ l, 5 mg/mL rPA5-AuNPs fluorescent optical sensor liquid are added, its fluorescence intensity are measured, due to rPA5-AuNPs solution energy
The fluorescence of quenching matter, therefore the fluorescence intensity of RhB solution reduces；RPA5-AuNPs fluorescent optical sensor liquid is continuously added, directly
Reach saturation (Fig. 6) to fluorescence intensity quenching.
Embodiment 5: cholesterol restores the fluorescence of RhB@rPA5-AuNPs
Cholesterol is configured to the liquid of 500 μm of ol/L with ethyl alcohol, adds 10 μ l, 2 μm of ol/L gallbladders solid every time into 4 solution of embodiment
Alcoholic solution, and static 3min fills it with RhB@rPA5-AuNPs in 4 solution of embodiment after addition cholesterol solution every time
Divide reaction, measure its fluorescence intensity, with the increase of cholesterol concentration, the fluorescence intensity of 4 solution of embodiment is gradually recovered (figure
7), and the linear relationship (Fig. 8) of fluorescence intensity and cholesterol concentration is obtained, for containing for the cholesterol in later period measurement sample
Amount；The range of linearity of cholesterol detection is 0.01-0.5 μM and 0.5-16 μM in the present embodiment, and detection is limited to 6.5 nM,
Equation of linear regression are as follows: the C of the C of F/F0=0.3488 (μM)+1.0131 and F/F0=0.0088 (μM)+1.1728,
Related coefficient is 0.935,0.991.
The interference free performance that embodiment 6:RhB@rPA5-AuNPs sensor identifies Cho
Cholesterol analog beta estradiol (β-Estradiol), estrone (Estrone) progress interference test are chosen, simultaneously
Also some conventional chaff interferents are chosen and carry out interference test, such as MgCl2, NaCl, KCl, glucose (Glucose), sucrose
(Sucrose), Tween-20, lysozyme (Lysozyme), dopamine (DA), glycine (Glycine), l-tyrosine (L-
Tyrosine).These chaff interferents are added to the mixed solution that rPA5-AuNPs solution quenching RhB fluorescence reaches saturation respectively
In (the fluorescence intensity quenching of embodiment 4 reaches the solution of saturation), its fluorescence recovery extent is measured, its corresponding fluorescence is made
Intensity (F-F0)/F0Image, see Figure 11, as can be seen from the figure RhB@rPA5-AuNPs sensor has Cho identification good
Interference free performance.
The use of embodiment 7:rPA5-AuNPs sensor
Using artificial serum as actual sample, the detection of cholesterol level in actual sample is carried out by the way of mark-on reclaims；It takes
50 times of 500 μ L of blood serum sample of dilution, distinguish in blood serum sample 500 μ L, 1 μm of ol/L of each addition, 2 μm of ol/L, 4 μm of ol/L,
The cholesterol solution of 8 μm of ol/L, it is respectively 0.5 μm of ol/L, 1 μm of ol/L, 2 μm of ol/L, 4 μm of ol/L that preparation, which obtains scalar quantity,
Sample to be tested will be added to 2mL, in 50mg/mL RhB@rPA5-AuNPs mixed liquor, measure each sample in sample to be tested
Fluorescent absorption value, each sample are measured in parallel 3 times, and it is strong that the fluorescent absorption value of each sample is substituted into the fluorescence that embodiment 5 obtains
Degree restores to calculate the actual measurement concentration of each sample, and further calculate recycling in the linear relationship with cholesterol concentration
Rate and standard deviation are shown in Table 1, and the experimental result display rate of recovery is between 95.3%-103.5%, relative standard deviation 2.8-
Between 4.5%, illustrate that sensor of the present invention can be used for detecting the cholesterol level in practical blood serum sample, biomedical and clinical
There is very big application potential in detection.
Cholesterol recovery testu result in 1 serum actual sample of table
。
Claims (5)
1. a kind of preparation method of fluorescent optical sensor, which is characterized in that steps are as follows:
(1) quinhydrones two (2- hydroxyethyl) ether, triphenylphosphine, anhydrous acetonitrile are sequentially placed into flask, it is cooling with ice-water bath, it stirs
After mixing mixing, carbon tetrabromide is added, is stirred to react at room temperature, cold water quenching reaction is added into mixture after complete reaction,
White precipitate is obtained, sediment is collected by filtration, is washed 3-4 times with methanol aqueous solution, with recrystallizing methanol, chemical combination is made after dry
Object 1, wherein the mass ratio of (2- hydroxyethyl) ether of quinhydrones two and triphenylphosphine is 0.1-1:1, quinhydrones two (2- hydroxyethyl) ether
Mass ratio with carbon tetrabromide is 0.1-0.5:1；
(2) compound 1, paraformaldehyde, 1,2- dichloroethanes are added in flask, and cooling with ice-water bath, then thereto
Boron trifluoride ether is added, is stirred to react at room temperature, after fully reacting plus water quenching is gone out, and is finally extracted with dichloromethane, organic phase
It is dry with anhydrous sodium sulfate, removing solvent is concentrated under reduced pressure and obtains crude product；Crude product obtains compound 2 by column chromatographic purifying,
Wherein the mass ratio of compound 1 and paraformaldehyde is 5-10:1, and compound 1 and the mass ratio of boron trifluoride ether are 0.5-2:
1；
(3) ethyl alcohol, compound 2,2- (dimethylamino) ethyl alcohol are added in flask, back flow reaction 24-72h, after reaction
It is cooled to room temperature, is concentrated under reduced pressure and removes solvent, residual solids are soluble in water, are filtered to remove insoluble matter, the rotation of gained filter vacuum
Solvent is evaporated off, finally by residue ethanol washing, vacuum drying obtains light tan solid column [5] aromatic hydrocarbons, wherein compound 2
Mass ratio with 2- (dimethylamino) ethyl alcohol is 0.1-0.5:1；
(4) PBS buffer solution, chlorauric acid solution, column [5] arene solution are added in water, is placed in mixed solution after stirring and evenly mixing
0.5-1.5h is reacted at 80-120 DEG C, solution is centrifuged after becoming claret, and precipitating is used deionized water eccentric cleaning 3-4 times, is made
RPA5-AuNPs fluorescent optical sensor, wherein the molar ratio of gold chloride and column [5] aromatic hydrocarbons is 0.01-0.05:1, water and PBS buffer solution
Volume ratio be 6-8:1, the volume ratio of PBS buffer solution and chlorauric acid solution is 5-6:1.
2. the preparation method of fluorescent optical sensor according to claim 1, it is characterised in that: methanol aqueous solution is first alcohol and water
3:2 is mixed to prepare by volume.
3. the preparation method of fluorescent optical sensor according to claim 1, it is characterised in that: PBS buffer solution 0.1mol/L,
The liquid of pH=7.0.
4. the preparation method of fluorescent optical sensor according to claim 1, it is characterised in that: the concentration of chlorauric acid solution is
0.01-0.02 mol/L, column [5] arene solution concentration are 0.02-0.04 mol/L.
5. the answering in cholesterol detection of fluorescent optical sensor made from the preparation method of fluorescent optical sensor described in claim 1
With.
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| CN109975260A (en) * | 2019-04-10 | 2019-07-05 | 山东大学 | A kind of method and its application based on nanogold fluorescence detection lysozyme |
| CN112816532A (en) * | 2020-12-24 | 2021-05-18 | 南通大学 | Photoelectric chemical sensor capable of regulating and controlling switch and preparation method and application thereof |
| CN114354582A (en) * | 2021-12-16 | 2022-04-15 | 江苏大学 | Preparation method of double-signal amplification electrochemiluminescence aptamer sensor and detection of Pb by using sensor2+Application of |
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| CN109975260A (en) * | 2019-04-10 | 2019-07-05 | 山东大学 | A kind of method and its application based on nanogold fluorescence detection lysozyme |
| CN112816532A (en) * | 2020-12-24 | 2021-05-18 | 南通大学 | Photoelectric chemical sensor capable of regulating and controlling switch and preparation method and application thereof |
| CN114354582A (en) * | 2021-12-16 | 2022-04-15 | 江苏大学 | Preparation method of double-signal amplification electrochemiluminescence aptamer sensor and detection of Pb by using sensor2+Application of |
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