CN112280556B - Preparation of phosphate radical responsive carbon quantum dots and application of phosphate radical responsive carbon quantum dots in fingerprint fluorescence identification - Google Patents
Preparation of phosphate radical responsive carbon quantum dots and application of phosphate radical responsive carbon quantum dots in fingerprint fluorescence identification Download PDFInfo
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Abstract
The preparation of the carbon quantum dots responding to the phosphate radical and the application of the carbon quantum dots in fingerprint fluorescence identification comprise the following steps; step 1: adding 1,2, 4-benzenetriol into ultrapure water and stirring to obtain a 1,2, 4-benzenetriol solution; and 2, step: putting the solution obtained in the step (1) into a high-pressure reaction kettle with a polytetrafluoroethylene lining for reaction; centrifuging after the reaction is finished, and taking supernatant for later use; and step 3: and (3) performing rotary evaporation and vacuum freeze drying on the supernatant obtained in the step (2) to obtain black solid powder, wherein the black solid powder is the carbon quantum dots. The invention has simple preparation process, low cost, good stability and no fluorescence in ultrapure water.
Description
Technical Field
The invention relates to the technical field of carbon quantum dot application, in particular to preparation of a phosphate radical responsive carbon quantum dot and application of the phosphate radical responsive carbon quantum dot in fingerprint fluorescence identification.
Background
Fingerprints have been widely used in the fields of forensic medicine, criminal investigation, biometrics, and medical diagnosis as important personal information, because fingerprints are unique to every person and do not change with age (Analytical methods, 2016, 8. In many cases, fingerprints are invisible (latent fingerprints), and they can only be seen directly with the naked eye after treatment with a specific method (Journal of Colloid and Interface Science,2018, 518. Therefore, developing new techniques for fingerprint "visualization" is crucial for the visual identification of invisible fingerprints. In general, sweat secreted from sweat pores on the finger surface contains various components such as phosphate (Pi), chloride, lipid, amino acid, etc., which pass through epidermal pores to reach the finger surface, and these secretions play an important role in the visualization or imaging of fingerprints (Analytical Chemistry,2019, 11185-11191. Currently, methods for visualizing imaged fingerprints are fluorescence, raman, fourier transform infrared, photoacoustic imaging, and the like. The fluorescence method is a microanalysis method with high sensitivity, good selectivity and low detection limit, overcomes the defects of complicated steps, long time consumption and the like of the traditional detection means, and can carry out qualitative and quantitative detection on the object to be detected in real time.
Carbon Quantum Dots (CQDs) have unique physicochemical properties, such as strong photoluminescence, good biocompatibility, easily modifiable structure, etc., and are therefore widely used in the fields of bioimaging, sensing, drug delivery, catalysis, energy conversion, cancer therapy, etc. The CQD is currently used for fingerprint imaging and mainly depends on electrostatic interaction between the CQD and finger sweat pore secretion components. This interaction is weak and not conducive to fine structure imaging of latent fingerprints. In addition, metal ions such as Cu can be used between CQD and finger sweat pore secretion component 2+ ，Fe 3+ ，Hg 2+ ，Fe 2+ Or Pb 2+ Interactive imaging fingerprinting is performed (Analytical and Bioanalytical Chemistry,2020, 412. The imaging method needs to be assisted by toxic metal ions, and meanwhile, optimization among multiple parameters is needed, so that the imaging method is not beneficial to large-scale generation and application. The CQD of the phosphoric acid (Pi) which is the main component secreted by the sweat pores of the fingers is selectively identified, the CQD for fluorescence imaging of the latent fingerprints is further developed based on the effect, and a new method for imaging the latent fingerprints is constructed, so that the method has important significance in detection, forensic science and criminal investigation of the phosphoric acid.
Disclosure of Invention
In order to overcome the technical problems, the invention aims to provide a preparation method of a carbon quantum dot responding to phosphate radicals and application of the carbon quantum dot to fingerprint fluorescence identification, the carbon quantum dot responding to phosphate radicals is prepared by a hydrothermal method, and the carbon quantum dot has the characteristics of rapidness, simplicity, convenience, high efficiency and low toxicity, is used as an enhanced fluorescent probe for qualitative detection and quantitative detection of phosphate radicals in a certain concentration range, and is applied to three-level (level 3) fluorescence imaging of latent fingerprints on paper by means of a mechanism that the carbon quantum dot and phosphoric acid directly act to enhance fluorescence.
In order to achieve the purpose, the invention adopts the technical scheme that:
a method for preparing a phosphate-responsive carbon quantum dot comprises the following steps;
step 1: adding 0.074-0.3mmol of 1,2, 4-benzenetriol into 50mL of ultrapure water, and stirring for 3 minutes to obtain a 1,2, 4-benzenetriol solution;
and 2, step: putting the solution obtained in the step (1) into a high-pressure reaction kettle with a polytetrafluoroethylene lining, and reacting for 4-8 hours at 150-180 ℃; centrifuging at 8000-12000 r/min after the reaction is finished, and taking supernatant liquor for later use;
and 3, step 3: and (3) performing rotary evaporation and vacuum freeze drying on the supernatant obtained in the step (2) to obtain black solid powder, wherein the black solid powder is the carbon quantum dots.
The structure of the 1,2, 4-benzenetriol is as follows:
a phosphate radical response carbon quantum dot is applied to a fingerprint fluorescence identification method, and the method comprises the following steps;
step 1: adding 0.3mg of the obtained carbon quantum dot powder into 1mL of deionized water to prepare 0.3mg/mL of mother solution for later use;
step 2: adding 200 mu L of the mother liquor obtained in the step 1 into 2.8mL of ultrapure water to prepare 20 mu g/mL of carbon quantum dot solution;
and 3, step 3: spraying the quantum dot solution obtained in the step (2) on a filter paper of 15cm multiplied by 15cm, and drying the filter paper at the temperature of 45-60 ℃ for later use;
and 4, step 4: pressing the finger on the filter paper obtained in the step 3, observing the filter paper under visible light after 5 seconds, imaging the filter paper without the fingerprint, and obviously observing a blue fluorescent fingerprint pattern under a 365nm ultraviolet lamp.
The carbon quantum dot mother solution obtained in the step 1 emits weak blue fluorescence, the maximum excitation wavelength of the carbon quantum dot mother solution is 370nm, the maximum emission peak is located at 464nm, the carbon quantum dot is 0.31-3.19 nm in size and has an average particle size of about 1.7 +/-0.38 nm when observed under a field emission transmission electron microscope, the spacing between crystal planes of the carbon quantum dot is 0.21nm when observed under a high-resolution transmission electron microscope, the carbon quantum dot prepared in an XRD (X-ray diffraction) spectrum has a characteristic peak with 26 degrees as a center corresponding to 100 crystal planes of graphite carbon, and the peak belongs to a (002) plane of graphitized carbon.
Respectively adding various ions and amino acids into the carbon quantum dot solution obtained in the step 2 for fluorescence test, wherein only PO is added 4 3- Then, the carbon quantum dot can emit strong blue fluorescence, the maximum excitation wavelength of the carbon quantum dot is 370nm, and the maximum emission peak is positioned at 448 nm.
The carbon quantum dot solution obtained in the step 2 is applied to qualitative detection and quantitative detection of phosphate radical in a certain concentration range (0-100 mu mol/L).
The quantum dot solution obtained in the step 2 is used for fluorescence imaging of fingerprints, after ordinary filter paper is sprayed and dried through carbon quantum dots, by means of a mechanism that fluorescence is enhanced through direct action of the carbon quantum dots and phosphoric acid, after the filter paper is pressed by a finger for 5s, an obvious blue fluorescence fingerprint pattern can be observed under 365nm ultraviolet light, and the fingerprint pattern comprises clear fingerprint details including level 1, level 2 and level 3.
The invention has the beneficial effects that:
the invention takes 1,2,4 benzenetriol as a carbon source and synthesizes the carbon quantum dots by a hydrothermal method. The carbon quantum dot has the advantages of simple preparation process, low cost, good stability and no fluorescence in ultrapure water.
Adding PO 4 3- After that, a significant blue fluorescence can be emitted immediately. The carbon quantum dot can be used as an enhanced fluorescent probe to selectively detect PO 4 3- . The carbon quantum dot has no obvious fluorescence change response to other common ions and amino acids, and can be used for PO in pure water 4 3- Qualitative and quantitative detection in a certain concentration range (0-100 mu mol/L). The carbon quantum dot pair PO 4 3- Has good detection effectAnd (4) sensitivity.
The carbon quantum dots synthesized by the method have weak autofluorescence on paper and only have the autofluorescence with PO 4 3- After the action, the blue fluorescence is obviously enhanced. The paper sprayed with the carbon quantum dot solution can be used for three-Level (Level 3) fluorescence imaging of fingerprints, and provides possibility for safety of paper documents.
Drawings
Fig. 1 is a particle size distribution diagram of the prepared carbon quantum dots.
Fig. 2 is a transmission electron micrograph of the prepared carbon quantum dot, and the inset is an enlarged crystal plane view.
Fig. 3 is an X-ray diffraction pattern of the prepared carbon quantum dots.
Fig. 4 is a graph of fluorescence excitation and emission spectra of the mother liquor of the prepared carbon quantum dots.
FIG. 5 is a graph of carbon quantum dots at a concentration of 20. Mu.g/mL in deionized water at a pH of 7.4 at a concentration of 260. Mu. Mol/L PO 4 3- There are fluorescence excitation spectra and emission spectra before and after.
FIG. 6 shows carbon quantum dots with 20. Mu.g/mL concentration in deionized water at pH 7.4 with PO concentrations of 0-180. Mu. Mol/L 4 3- Graph of the change in fluorescence intensity at 448nm thereafter.
FIG. 7 shows carbon quantum dots at a concentration of 20. Mu.g/mL in deionized water at pH 7.4 with PO added at a concentration of 0-180. Mu. Mol/L 4 3- Logarithmic fluorescence intensity values (base 10) at 448nm after 4 3- Linear dependence of concentration.
FIG. 8 is a graph of fluorescence intensity of carbon quantum dots at a concentration of 20. Mu.g/mL in deionized water at pH 7.4 at 448nm after addition of common cations at a concentration of 140. Mu. Mol/L, respectively.
FIG. 9 is a bar graph of fluorescence intensity at 448nm of carbon quantum dots at a concentration of 20 μ g/mL in deionized water at pH 7.4 after addition of common cations at a concentration of 140 μmol/L, respectively.
FIG. 10 is a graph of fluorescence intensity at 448nm of carbon quantum dots at a concentration of 20. Mu.g/mL in deionized water at pH 7.4 after addition of common anions at a concentration of 140. Mu. Mol/L, respectively.
FIG. 11 is a bar graph of fluorescence intensity at 448nm of carbon quantum dots at a concentration of 20 μ g/mL in deionized water at a pH of 7.4 after addition of a concentration of 266.67 μmol/L of common anions, respectively.
FIG. 12 is a graph of fluorescence intensity at 448nm of carbon quantum dots at a concentration of 20. Mu.g/mL in deionized water at pH 7.4 after addition of common biomolecules at a concentration of 140. Mu. Mol/L, respectively.
FIG. 13 is a bar graph of fluorescence intensity at 448nm of carbon quantum dots at a concentration of 20 μ g/mL in deionized water at pH 7.4 after addition of common biomolecules at a concentration of 140 μmol/L, respectively.
FIG. 14 is a fluorescent fingerprint pattern and fingerprint detail including level 1, level 2 and level 3 under 365nm ultraviolet light after a filter paper sprayed with carbon quantum dots is pressed by a finger for 5 seconds, and the scale is 1cm.
Detailed Description
The present invention will be described in further detail with reference to examples.
Preparation of phosphate-responsive carbon quantum dots.
Example 1:
1. 50mL of ultrapure water was added to a 100mL beaker, followed by 0.15mmol of 1,2, 4-benzenetriol, and the mixture was magnetically stirred for 3 minutes to obtain a 1,2, 4-benzenetriol solution.
2. And (2) putting the 1,2, 4-benzenetriol solution obtained in the step (1) into a high-pressure reaction kettle with a polytetrafluoroethylene lining, reacting for 6 hours at 170 ℃, centrifuging at 8000r/min after the reaction is finished, and taking supernatant for later use.
3. And (3) performing rotary evaporation and vacuum freeze drying on the supernatant obtained in the step (2) to obtain black solid powder.
4. 0.3mg of the solid powder obtained in step 3 was dissolved in 1mL of ultrapure water to obtain a mother liquor for use.
5. Observing the mother liquor under a field emission transmission electron microscope, wherein the carbon quantum dots have the size of 0.31-3.19 nm and the average particle size of about 1.7 +/-0.38 nm (see attached figure 1). The obtained solid powder is tested by a high-resolution transmission electron microscope, the particles are nearly spherical, the interplanar spacing of carbon quantum dots is 0.21nm (shown in figure 2), and the interplanar spacing belongs to 100 crystal planes of graphitized carbon respectively.
6. The obtained solid powder was subjected to XRD measurement, and the synthesized powder had a characteristic peak centered at 26 ° (see fig. 3), which belongs to the (002) plane of graphitized carbon. The solid powder is carbon quantum dot powder.
7. And (3) performing fluorescence performance test on the carbon quantum dot mother liquor obtained in the step (4), wherein the carbon quantum dot mother liquor has weaker fluorescence, the maximum excitation wavelength is 370nm, and the maximum emission wavelength is 464nm (see the attached figure 4).
Example 2:
1. 50mL of ultrapure water was added to a 100mL beaker, followed by 0.074mmol of 1,2, 4-benzenetriol, and the mixture was magnetically stirred for 3 minutes to obtain a 1,2, 4-benzenetriol solution.
2. And (2) putting the 1,2, 4-benzenetriol solution obtained in the step (1) into a high-pressure reaction kettle with a polytetrafluoroethylene lining, reacting for 4 hours at 150 ℃, centrifuging at 8000r/min after the reaction is finished, and taking supernatant for later use.
3. And (3) performing rotary evaporation and vacuum freeze drying on the supernatant obtained in the step (2) to obtain black solid powder.
4. 0.3mg of the solid powder obtained in step 3 was dissolved in 1mL of ultrapure water to obtain a mother liquor for use.
Example 3:
1. 50mL of ultrapure water was added to a 100mL beaker, followed by 0.3mmol of 1,2, 4-benzenetriol, and the mixture was magnetically stirred for 3 minutes to obtain a 1,2, 4-benzenetriol solution.
2. And (2) putting the 1,2, 4-benzenetriol solution obtained in the step (1) into a high-pressure reaction kettle with a polytetrafluoroethylene lining, reacting for 8 hours at 180 ℃, centrifuging at 12000r/min after the reaction is finished, and taking supernatant for later use.
3. And (3) performing rotary evaporation and vacuum freeze drying on the supernatant obtained in the step (2) to obtain black solid powder.
4. 0.3mg of the solid powder obtained in step 3 was dissolved in 1mL of ultrapure water to obtain a mother liquor for use.
The prepared carbon quantum dots are applied to qualitative and quantitative detection of phosphate radicals.
1. 200 μ L of 0.3mg/mL of the mother liquor was added to 2.8mL of ultrapure water, and 260 μmol/L of PO was added 4 3- The fluorescence performance test was carried out with a maximum excitation wavelength of 370nm and a maximum emission wavelength of 448nm (see FIG. 5).
2. 0.04mg of the carbon quantum dot solid powder was added to 3mL of ultrapure water (pH = 7.4), and PO was added to the solution at a concentration of 0, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 149, 150, 160, 170, 180 μmol/L 4 3- The fluorescence spectrum was detected (see FIG. 6). Adding 20-180 mu mol/L PO 4 3- The logarithmic value (base 10) of the fluorescence intensity at the emission wavelength of 448nm of the obtained product is determined according to different PO 4 3- The concentration has good linear relation, the fitting equation is y =0.01015x-3.1757, and the correlation coefficient R 2 =0.996, detection limit 0.34 μmol/L (see fig. 7).
3. 0.04mg of the carbon quantum dot solid powder was added to 3mL of ultrapure water (pH = 7.4), and Ag was added to the solution at a concentration of 140. Mu. Mol/L + ，Ba 2+ ，Ca 2+ ，Cd 2+ ，Cr 3+ ，Cu 2+ ，Fe 2+ ，Fe 3+ ，Hg 2+ ，K + ，Mg 2+ ，Mn 2+ ，Ni 2+ And Zn 2+ And the change in fluorescence intensity was detected (see FIG. 8). Only adding PO 4 3- Blue fluorescence was detected at 448nm without response from the addition of these common cationic species. The relative fluorescence intensity change at 448nm was observed by adding different cations, respectively (FIG. 9).
4. 0.04mg of the carbon quantum dot solid powder was added to 3mL of ultrapure water (pH = 7.4), and F was added to the solution thereof at a concentration of 140 μmol/L, respectively - ，Cl - ，Br - ，CO 3 2- ，HCO 3 - ，I - ，NO 3 - ，SO 3 2- And PO 4 3- And the change in fluorescence intensity was detected (see FIG. 10). Only adding PO 4 3- Blue fluorescence was detected at 448nm without response by the addition of these common anionic species. The relative fluorescence intensity change at 448nm was observed after addition of different anions (FIG. 11).
5. 0.04mg of the carbon quantum dot solid powder was added to 3mL of ultrapure water (pH = 7.4), and L-phenylalanine, L-alanine, DL-methionine, glycine, leucine, tryptophan, L-threonine, valine, histidine, homocysteine, glucose and cysteine were added to the solution thereof at a concentration of 140. Mu. Mol/L, respectively, and the change in fluorescence intensity was detected (see FIG. 12). Only adding PO 4 3- Blue fluorescence was detected at 448nm, but no response was observed by addition of these common amino acid species. The relative change in fluorescence intensity at 448nm was observed after addition of different amino acids, respectively (FIG. 13).
The prepared carbon quantum dots are applied to the fluorescent identification of fingerprints on paper.
1. 0.08mg of the carbon quantum dot solid powder was added to 3mL of ultrapure water (pH = 7.4) to obtain a carbon quantum dot solution of 26.6 μ g/mL.
2. The solution obtained was sprayed onto a filter paper of 15cm × 15cm and the filter paper was dried in an oven at 45 ℃. Press with a finger on the filter paper for 5 seconds. Under natural light, no fingerprint on the filter paper was observed. A blue fluorescent fingerprint pattern was clearly observed under 365nm uv light. The fingerprint pattern comprises clear fingerprint details including fingerprint outlines, threads (level 1), short ridges, ridge bifurcations (level 2) and sweat pores (level 3) (see attached figure 14).
Claims (9)
1. A method for preparing a phosphate-responsive carbon quantum dot, comprising the steps of;
step 1: adding 0.074 to 0.3mmol of 1,2, 4-benzenetriol into 50mL of ultrapure water, and stirring to obtain a 1,2, 4-benzenetriol solution;
and 2, step: putting the solution obtained in the step 1 into a high-pressure reaction kettle with a polytetrafluoroethylene lining for reaction; centrifuging after the reaction is finished, and taking supernatant for later use;
and step 3: and (3) performing rotary evaporation and vacuum freeze drying on the supernatant obtained in the step (2) to obtain black solid powder, wherein the black solid powder is the carbon quantum dots.
2. The method for preparing a phosphate-responsive carbon quantum dot according to claim 1, wherein the stirring of step 1 is performed for 3 minutes.
3. The method for preparing the carbon quantum dot responding to the phosphate radical according to claim 1, wherein the reaction is carried out in the high-pressure reaction kettle of the step 2 at 150 to 180 ℃ for 4 to 8 hours.
4. The method for preparing the phosphate radical responsive carbon quantum dot according to claim 1, wherein the step 2 is centrifugal at a rotating speed of 8000-12000 r/min after the high-temperature reaction.
5. The carbon quantum dot prepared by the method of claim 1 is applied to a fingerprint fluorescence identification method, and comprises the following steps;
step 1: adding 0.3mg of the obtained carbon quantum dot powder into 1mL of deionized water to prepare 0.3mg/mL of mother solution for later use;
step 2: adding 200 mu L of the mother liquor obtained in the step 1 into 2.8mL of ultrapure water to prepare a carbon quantum dot solution of 20 mu g/mL;
and step 3: spraying the carbon quantum dot solution obtained in the step (2) on 15 cm-15 cm filter paper, and drying the filter paper at the temperature of 45-60 ℃ for later use;
and 4, step 4: pressing the finger on the filter paper obtained in the step 3, observing the filter paper under visible light after 5 seconds, imaging the filter paper without the fingerprint, and obviously observing a blue fluorescent fingerprint pattern under a 365nm ultraviolet lamp.
6. The method for fluorescence fingerprint identification by using carbon quantum dots as claimed in claim 5, wherein the quantum dot solution obtained in step 2 is used for fluorescence imaging of fingerprints, after ordinary filter paper is sprayed and dried by the carbon quantum dots, and by means of the mechanism that fluorescence is enhanced by direct action of the carbon quantum dots and phosphoric acid, after the filter paper is pressed by fingers for 5s, obvious blue fluorescence fingerprint patterns including fingerprint details including clear fingerprint outlines, threads, ridge bifurcations and sweat pores can be observed under 365nm ultraviolet light.
7. The method for fingerprint fluorescence identification of carbon quantum dots according to claim 5, wherein the carbon quantum dot mother liquor obtained in step 1 emits weak blue fluorescence, the maximum excitation wavelength is 370nm, the maximum emission peak is at 464nm, the size of the carbon quantum dot is 0.31 to 3.19nm and the average particle size is 1.7 ± 0.38nm when observed by a field emission transmission electron microscope, the spacing between crystal planes of the carbon quantum dot is 0.21nm when observed by a high-resolution transmission electron microscope, the carbon quantum dot prepared in the XRD spectrum corresponds to 100 crystal planes of graphite carbon, and the peak belongs to (002) plane of graphitized carbon.
8. The method for detecting fluorescence intensity of carbon quantum dots is characterized in that 0.3mg of the carbon quantum dot powder is added into 1mL of deionized water to prepare 0.3mg/mL of mother solution for later use; adding 200 mu L of the obtained mother liquor into 2.8mL of ultrapure water to prepare a 20 mu g/mL carbon quantum dot solution; the carbon quantum dot solution is applied to qualitative detection and quantitative detection of phosphate radical within the concentration range of 0 to 100 mu mol/L, and the detection limit is 0.34 mu mol/L.
9. The application of the carbon quantum dots in the detection of fluorescence intensity of the carbon quantum dots according to claim 8, wherein various ions and amino acids are respectively added into the carbon quantum dot solution for fluorescence test, and only PO is added 4 3- Then, the carbon quantum dot can emit strong blue fluorescence, the maximum excitation wavelength of the carbon quantum dot is 370nm, and the maximum emission peak is at 448 nm.
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