CN113548657A - Natural plant impatiens carbon dots and preparation method and application thereof - Google Patents
Natural plant impatiens carbon dots and preparation method and application thereof Download PDFInfo
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Abstract
The invention relates to the field of fluorescent nano material application, and particularly discloses a natural plant impatiens carbon dot and a preparation method and application thereof. The fluorescence performance of the carbon dots of the impatiens balsamina is evaluated on a fluorometer, and the result shows that the quantum yield of the carbon dots of the impatiens balsamina is 23%, the carbon dots have excellent fluorescence property, can selectively image gram-positive bacteria so as to identify the gram-positive bacteria and the gram-negative bacteria, and the carbon dots of the impatiens balsamina have good water solubility and low cytotoxicity and have a bacteriostatic effect on the gram-positive bacteria.
Description
Technical Field
The invention belongs to the field of fluorescent nano material application, and particularly relates to a natural plant impatiens carbon dot and a preparation method and application thereof.
Background
Bacterial infections have become one of the leading causes of death worldwide and are a global challenge to human health. Although antibiotic therapy has been widely used in recent years with the development of medical technology, the development of new antibacterial agents for clinical treatment is imminent due to the increasing number of bacterial infections caused by the gradual emergence of antibiotic resistance. In addition, before antibiotic treatment is selected, it is important to distinguish whether the pathogenic bacteria are gram-positive bacteria or gram-negative bacteria for clinical treatment. The method not only can preliminarily identify the bacteria, but also has different action mechanisms of gram-positive bacteria and gram-negative bacteria, has different sensitivity degrees to different antibiotics, and can provide a basis for treatment by identifying the types of pathogenic bacteria. Although gram-positive bacteria and gram-negative bacteria can be identified by the traditional gram staining method, errors are easy to occur due to the fact that the gram-positive bacteria and the gram-negative bacteria are observed by naked eyes under a microscope, and therefore the development of a more accurate and convenient method for distinguishing the gram-positive bacteria and the gram-negative bacteria is urgently needed.
With the development of polymer materials in recent years, nano materials are also considered to have good antibacterial performance, and various nano materials containing metal elements, such as gold nanoparticles, silver nanoparticles, titanium dioxide nanoparticles and the like, have been successfully developed and have high antibacterial performance, but the potential cytotoxicity and long-term in vivo retention of the nano materials still have certain challenges in clinical use. Carbon dots have been widely used in the fields of sensing, bio-imaging, drug delivery, photocatalysis, etc. due to their adjustable size, surface charge, low cytotoxicity, good biocompatibility, and cell membrane damaging properties.
The impatiens balsamina is a traditional Chinese medicinal material, and the extracts of the impatiens balsamina in different solvents have been found to have good antioxidant and antibacterial properties in the prior research. The eutectic solvent (DES) is a new green solvent, and has the advantages of low cost, no toxicity, wide dissolving range and good biocompatibility. And multiple researches find that the doping of heteroatoms such as nitrogen, sulfur, boron, fluorine, chlorine and the like in the carbon dots can effectively improve the quantum yield, the oxidation resistance and the antibacterial performance of the carbon dots. Whereas DES contains a large number of heteroatoms necessary for doping carbon sites.
Disclosure of Invention
The invention aims to provide a natural plant impatiens carbon dot which has a quantum yield of 23%, has excellent fluorescence property, can selectively image gram-positive bacteria so as to identify the gram-positive bacteria and the gram-negative bacteria and has an antibacterial effect.
The invention provides a preparation method of natural plant impatiens balsamina carbon dots, which comprises the following steps:
step 1, adding impatiens balsamina powder into a solvent, performing ultrasonic treatment, and reacting in a hydrothermal reaction kettle for 5-10h at the reaction temperature of 200-280 ℃;
the solvent is a mixed solution of a eutectic solvent and pure water;
step 2, after cooling to room temperature, centrifuging at 10000rpm of 8000-.
Further, in the step 1, the eutectic solvent consists of choline chloride and acetamide according to a molar ratio of 1:1-2, or the choline chloride and the urea are mixed according to the mol ratio of 1:1-2, or the choline chloride and the malonic acid are mixed according to the mol ratio of 1:1-2, or the choline chloride and the acetic acid according to the molar ratio of 1:1-2, or the choline chloride and the formic acid according to the molar ratio of 1:1-2, or choline chloride and malic acid according to the molar ratio of 1:1-2, or tetrabutylammonium bromide and glycerol according to the molar ratio of 1-2:1-3, or the tetrabutylammonium bromide and the ethylene glycol in a molar ratio of 1:1-2, or the tetrabutylammonium bromide and the acetic acid in a molar ratio of 1:1-2, or the tetrabutylammonium bromide and the malonic acid in a molar ratio of 1: 1-2.
Further, the eutectic solvent consists of choline chloride and acetamide in a molar ratio of 1: 1.
Further, in step 1, the impatiens balsamina powder is obtained by drying and pulverizing impatiens balsamina petals and sieving.
Further, in the step 1, the ultrasonic time is 10-30 min.
Further, in the step 1, the mass ratio of the impatiens balsamina powder to the solvent is 1: 10-30.
Further, in the step 1, pure water accounts for 0-30% of the volume of the solvent.
The invention also provides the natural plant impatiens balsamina carbon dots prepared by the preparation method.
The invention also provides application of the carbon dots of the natural plant impatiens balsamina in gram-positive bacteria identification.
The invention also provides application of the natural plant impatiens balsamina carbon dots in antibiosis.
Compared with the prior art, the natural plant impatiens carbon dots and the preparation method and the application thereof provided by the invention have the following beneficial effects:
1. the preparation process of the natural plant impatiens balsamina carbon dots is simple, and the preparation cost is low.
2. The carbon dots of the natural plant impatiens balsamina have excellent fluorescence performance, the quantum yield is high and reaches 23%, gram-positive bacteria can be specifically identified through multicolor fluorescence imaging (blue, green and red) of the gram-positive bacteria, and gram-negative bacteria cannot be imaged, so that the identification of the gram-positive bacteria and the gram-negative bacteria is carried out.
3. The natural plant impatiens balsamina carbon dots prepared by the method can be used as an antibacterial agent, can inhibit the growth of gram-positive bacteria at low concentration and can inhibit the growth of gram-negative bacteria at high concentration, so that the method can be used for pertinently preventing and treating diseases caused by the gram-positive bacteria.
4. The bacteriostatic mechanism of the carbon dots of the natural plant impatiens balsamina prepared by the invention is mainly to generate free radicals, thereby ensuring the application potential of the bacteriostatic agent to drug-resistant bacteria.
5. The natural plant impatiens balsamina carbon dot prepared by the method has low cytotoxicity, and MTT (methyl thiazolyl tetrazolium) results of two normal cells (keratinocytes and macrophages) and two cancer cells (liver cancer cells and breast cancer cells) show that the cells still have about 80% survival rate under the condition that the concentration of the impatiens balsamina carbon dot is 3mg/mL, so that the impatiens balsamina carbon dot has the potential of being applied in vivo as a bacteriostatic agent.
6. According to the invention, the impatiens balsamina is used as a carbon source, and the eutectic solvent is used as a solvent and provides a doping element, so that the fluorescent carbon dots with excellent antibacterial property are finally prepared. The invention modifies a large amount of amino on the surface of the carbon point, changes the surface potential of the carbon point and greatly increases the antibacterial performance of the carbon point. Meanwhile, the negative charge on the surface of gram-positive bacteria is far more than that of gram-negative bacteria, so that the bacteriostatic effect of the gram-positive bacteria on the gram-positive bacteria is far more than that of the gram-negative bacteria. In addition, the fluorescent carbon dots have high quantum yield and positive charges on the surface, so that the fluorescent carbon dots can kill bacteria and selectively image gram-positive bacteria. Therefore, the impatiens balsamina carbon dots obtained by the method can effectively distinguish gram-positive bacteria from gram-negative bacteria.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.
FIG. 1 is a schematic flow chart of the preparation of carbon dots of the natural plant impatiens balsamina in example 1 of the present invention;
FIG. 2 is an electron micrograph and a particle size distribution analysis of carbon dots of the natural plant impatiens balsamina prepared in example 1 of the present invention;
wherein, the figure a shows a transmission electron microscope image of carbon dots (N, Cl-CDs) of the natural plant impatiens balsamina prepared in example 1 of the present invention;
FIG. b is a high-resolution TEM image of carbon dots (N, Cl-CDs) of the natural plant balsam prepared in example 1 of the present invention;
FIG. c is a graph showing the analysis of the particle size distribution of the natural plant impatiens balsamina carbon dots (N, Cl-CDs) prepared in example 1 of the present invention, wherein the abscissa shows the length of the diameter and the ordinate shows the distribution;
FIG. 3 is a graph showing the gram-positive bacteria inhibition test effect of carbon dots (N, Cl-CDs) of the natural plant impatiens balsamina prepared in example 1 of the present invention;
wherein, the graph a shows the influence of carbon points (N, Cl-CDs) of the natural plant impatiens balsamina on the OD value of staphylococcus aureus in different concentrations;
FIG. b is a graph showing the effect of different concentrations of carbon dots (N, Cl-CDs) of the natural plant impatiens balsamina on the OD value of enterococcus faecium;
FIG. c is a graph showing the effect of different concentrations of carbon dots (N, Cl-CDs) of the natural plant impatiens balsamina on the OD value of Bacillus subtilis;
FIG. 4 shows MTT results of natural plant impatiens balsamina carbon dots (N, Cl-CDs) versus different cells prepared in example 1 of the present invention;
wherein, the graph a shows the influence of carbon dots (N, Cl-CDs) of the natural plant impatiens balsamina with different concentrations on the vitality of liver cancer cells;
panel b is a graph of the effect of different concentrations of the carbon dots (N, Cl-CDs) of the natural plant impatiens balsamina on breast cancer cell viability;
FIG. c is a graph of the effect of different concentrations of carbon dots (N, Cl-CDs) of the natural plant impatiens balsamina on keratinocyte viability;
FIG. d is a graph of the effect of different concentrations of carbon dots (N, Cl-CDs) of the natural plant impatiens balsamina on macrophage viability;
FIG. 5 is a fluorescent inverted microscope photograph of gram-positive bacteria incubated with carbon dots (N, Cl-CDs) of the natural plant impatiens balsamina prepared in example 1 of the present invention for 2 hours, wherein S.aureus represents Staphylococcus aureus, B.subtilis represents Bacillus subtilis, and E faecium represents enterococcus faecium;
FIG. 6 is a fluorescent inverted microscope photograph of the natural plant impatiens balsamina prepared in example 1 of the present invention, in which E coli represents E.coli, P.aeruginosa represents Pseudomonas aeruginosa, and Salmonella represents Salmonella, incubated with gram-negative bacteria for 2 hours.
Detailed Description
The present invention is described in detail below with reference to the drawings and the specific embodiments, but it should be understood that the scope of the present invention is not limited by the specific embodiments. The test methods in the following examples, which are not specified in specific conditions, are generally conducted under conventional conditions, and the steps thereof will not be described in detail since they do not relate to the invention.
When numerical ranges are given in the examples, it is understood that both endpoints of each of the numerical ranges and any value therebetween can be selected unless the invention otherwise indicated. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In addition to the specific methods, devices, and materials used in the examples, any methods, devices, and materials similar or equivalent to those described in the examples may be used in the practice of the invention in addition to the specific methods, devices, and materials used in the examples, in keeping with the knowledge of one skilled in the art and with the description of the invention.
Preparation of carbon dots of impatiens balsamina L.var.balsamina L.
Example 1
A preparation method of natural plant impatiens balsamina carbon dots specifically comprises the following steps:
step 1, drying and crushing impatiens balsamina petals at 60 ℃, and then sieving the impatiens balsamina petals with a 40-mesh sieve to obtain impatiens balsamina powder, wherein the mass ratio of the impatiens balsamina powder is 1: 20, adding the mixture into a solvent, performing ultrasonic treatment for 30min, and reacting in a hydrothermal reaction kettle at 240 ℃ for 7.5 hours to form a carbon dot solution;
the solvent is a eutectic solvent, namely does not contain pure water;
the eutectic solvent is prepared by taking choline chloride as a hydrogen ion receptor and acetamide as a hydrogen ion donor, wherein the molar ratio of the choline chloride to the acetamide is 1: 1;
step 2, after cooling to room temperature (25 ℃), centrifuging at 9000rpm for 20min to remove precipitates, dialyzing the supernatant in ultrapure water for 24 hours by using a dialysis bag with molecular weight cutoff of 1000, carrying out rotary evaporation on the dialyzed solution at 80 ℃ to obtain a pure carbon dot solution, carrying out vacuum freeze drying at-20 ℃ to obtain purified natural plant impatiens balsamina carbon dot powder, wherein the particle size characterization result is shown in figure 2, and the preparation flow is shown in figure 1.
Example 2
This example provides a method for preparing carbon dots from impatiens balsamina, which comprises the following steps:
in the step 1, the mass ratio of the impatiens balsamina powder to the solvent is 1:10, performing ultrasonic treatment for 10min, and reacting in a hydrothermal reaction kettle at 200 ℃ for 10 hours;
the volume fraction of pure water in the solvent is 30%;
in step 2, the centrifugation condition is 10000rpm for 10min, a 600Da dialysis bag is used for dialysis for 72h, and the rotary evaporation temperature is 100 ℃.
Example 3
This example provides a method for preparing touch-me-not carbon dots, which includes the steps substantially the same as those of example 1, except that:
in the step 1, the mass ratio of the impatiens balsamina powder to the solvent is 1: 30, carrying out ultrasonic treatment for 20min, and reacting for 5 hours at 280 ℃ in a hydrothermal reaction kettle;
in the solvent, the volume fraction of pure water is 10%;
in the step 2, the centrifugation condition is 8000rpm for 30min, the dialysis is carried out for 48h by using a 800Da dialysis bag, and the rotary evaporation temperature is 60 ℃.
Example 4
This example provides a method for preparing touch-me-not carbon dots, which includes the steps substantially the same as those of example 1, except that:
in the step 1, the mass ratio of the impatiens balsamina powder to the solvent is 1: 25, performing ultrasonic treatment for 25min, and reacting for 5 hours at 250 ℃ in a hydrothermal reaction kettle;
the volume fraction of pure water in the solvent was 20%.
Example 5
This example provides a method for preparing touch-me-not carbon dots, which includes the steps substantially the same as those of example 1, except that:
the eutectic solvent consists of choline chloride and acetamide according to a molar ratio of 1: 2.
Example 6
This example provides a method for preparing touch-me-not carbon dots, which includes the steps substantially the same as those of example 1, except that:
the eutectic solvent consists of choline chloride and urea according to a molar ratio of 1: 1.
Example 7
This example provides a method for preparing touch-me-not carbon dots, which includes the steps substantially the same as those of example 1, except that:
the eutectic solvent consists of choline chloride and urea according to a molar ratio of 1: 2.
Example 8
This example provides a method for preparing touch-me-not carbon dots, which includes the steps substantially the same as those of example 1, except that:
the eutectic solvent consists of choline chloride and malonic acid according to a molar ratio of 1: 1.
Example 9
This example provides a method for preparing touch-me-not carbon dots, which includes the steps substantially the same as those of example 1, except that:
the eutectic solvent consists of choline chloride and malonic acid according to a molar ratio of 1: 2.
Example 10
This example provides a method for preparing touch-me-not carbon dots, which includes the steps substantially the same as those of example 1, except that:
the eutectic solvent consists of choline chloride and acetic acid according to a molar ratio of 1: 1.
Example 11
This example provides a method for preparing touch-me-not carbon dots, which includes the steps substantially the same as those of example 1, except that:
the eutectic solvent consists of choline chloride and acetic acid according to a molar ratio of 1: 2.
Example 12
This example provides a method for preparing touch-me-not carbon dots, which includes the steps substantially the same as those of example 1, except that:
the eutectic solvent consists of choline chloride and formic acid according to a molar ratio of 1: 1.
Example 13
This example provides a method for preparing touch-me-not carbon dots, which includes the steps substantially the same as those of example 1, except that:
the eutectic solvent consists of choline chloride and formic acid according to a molar ratio of 1: 2.
Example 14
This example provides a method for preparing touch-me-not carbon dots, which includes the steps substantially the same as those of example 1, except that:
the eutectic solvent consists of choline chloride and malic acid according to a molar ratio of 1: 1.
Example 15
This example provides a method for preparing touch-me-not carbon dots, which includes the steps substantially the same as those of example 1, except that:
the eutectic solvent consists of choline chloride and malic acid according to a molar ratio of 1: 2.
Example 16
This example provides a method for preparing touch-me-not carbon dots, which includes the steps substantially the same as those of example 1, except that:
the eutectic solvent consists of tetrabutylammonium bromide and glycerol according to the molar ratio of 1: 1.
Example 17
This example provides a method for preparing touch-me-not carbon dots, which includes the steps substantially the same as those of example 1, except that:
the eutectic solvent consists of tetrabutylammonium bromide and glycerol according to the molar ratio of 1: 3.
Example 18
This example provides a method for preparing touch-me-not carbon dots, which includes the steps substantially the same as those of example 1, except that:
the eutectic solvent consists of tetrabutylammonium bromide and glycerol according to the molar ratio of 2: 1.
Example 19
This example provides a method for preparing touch-me-not carbon dots, which includes the steps substantially the same as those of example 1, except that:
the eutectic solvent consists of tetrabutylammonium bromide and ethylene glycol according to a molar ratio of 1: 1.
Example 20
This example provides a method for preparing touch-me-not carbon dots, which includes the steps substantially the same as those of example 1, except that:
the eutectic solvent consists of tetrabutylammonium bromide and ethylene glycol according to a molar ratio of 1: 2.
Example 21
This example provides a method for preparing touch-me-not carbon dots, which includes the steps substantially the same as those of example 1, except that:
the eutectic solvent consists of tetrabutylammonium bromide and acetic acid according to a molar ratio of 1: 1.
Example 22
This example provides a method for preparing touch-me-not carbon dots, which includes the steps substantially the same as those of example 1, except that:
the eutectic solvent consists of tetrabutylammonium bromide and acetic acid according to a molar ratio of 1: 2.
Example 23
This example provides a method for preparing touch-me-not carbon dots, which includes the steps substantially the same as those of example 1, except that:
the eutectic solvent consists of tetrabutylammonium bromide and malonic acid according to a molar ratio of 1: 1.
Example 24
This example provides a method for preparing touch-me-not carbon dots, which includes the steps substantially the same as those of example 1, except that:
the eutectic solvent consists of tetrabutylammonium bromide and malonic acid according to a molar ratio of 1: 2.
The properties of the carbon dots of the impatiens balsamina prepared in the embodiments 1 to 24 of the present invention are similar, and the best embodiment 1 is taken as a representative to carry out bacteriostasis experimental study, specifically as follows:
secondly, study on inhibition of carbon points of the natural plant impatiens balsamina prepared in the embodiment 1 of the invention on gram-positive bacteria
1. The inhibition effect of the obtained natural plant impatiens balsamina carbon dot on staphylococcus aureus prepared in example 1 is as follows:
selecting Staphylococcus aureus in exponential growth phase, and adjusting its concentration to OD6000.5, then mixing with 1: 100 was diluted with carbon dots (0.00, 0.02, 0.04, 0.06, 0.08, 0.10mg/mL) containing various concentrations of impatiens balsamina. Culturing in an incubator at 37 deg.C for 24 hr, wherein the bacterial solution has no obvious turbidity and no obvious increase in turbidity at 600nm, and the concentration of the drug added is MIC of carbon dots of impatiens balsamina to Staphylococcus aureus.
The results are shown in FIG. 3a, where the MIC of the carbon dot of impatiens balsamina to Staphylococcus aureus is 0.08 mg/mL.
2. The inhibition effect of the obtained natural plant impatiens balsamina carbon dots on enterococcus faecium prepared in example 1 is as follows:
selecting enterococcus faecium in exponential growth phase, and adjusting its concentration to OD6000.5, then mixing with 1: 100 was diluted with carbon dots (0.00, 0.05, 0.10, 0.15, 0.20mg/mL) containing various concentrations of impatiens balsamina. Culturing in a 37 ℃ incubator for 24 hours, wherein the bacterial liquid has no obvious turbidity and the turbidity at 600nm is not obviously increased, and the adding concentration is the MIC of the impatiens carbon dots to the enterococcus faecium. The results are shown in FIG. 3b, where the MIC of the C point of impatiens balsamina to enterococcus faecium is 0.2 mg/mL.
3. The inhibition effect of the obtained natural plant impatiens balsamina carbon dot on bacillus subtilis prepared in example 1 is specifically as follows:
selecting Bacillus subtilis in exponential growth phase, and adjusting its concentration to OD6000.5, then mixing with 1: 100 was diluted with carbon dots (0.00, 0.04, 0.08, 0.12, 0.16mg/mL) containing different concentrations of impatiens balsamina. Culturing in an incubator at 37 ℃ for 24 hours, wherein the bacterial liquid is not obviously turbid, the turbidity at 600nm is not obviously increased, and the adding concentration is the MIC of the impatiens carbon dots to the bacillus subtilis. The results are shown in FIG. 3c, where the MIC of the carbon point of impatiens balsamina to Bacillus subtilis is 0.16 mg/mL.
Thirdly, study on carbon point cytotoxicity of the natural plant impatiens balsamina prepared in example 1 of the present invention
1. The cytotoxicity of the carbon point of the natural plant impatiens balsamina prepared in the example 1 on liver cancer cells is studied, and the specific method is as follows:
hepatoma cells (HepG2) were selected, and the toxicity of carbon dots at concentrations of 0, 0.1, 0.2, 0.4, 0.6, 0.8, 1.0, 2.0, 3.0mg/mL (measured 24 hours after the addition of carbon dots to the cells) to HepG2 was measured by MTT assay using a microplate reader, and the results are shown in FIG. 4 a. The experimental result shows that the survival rate of cells still exceeds 90% under the condition that the concentration of the carbon dots of the impatiens balsamina is far greater than the Minimum Inhibitory Concentration (MIC) of gram-positive bacteria, which indicates that the carbon dots of the impatiens balsamina have good biocompatibility.
2. The cytotoxicity of the obtained natural plant impatiens balsamina carbon dot prepared in example 1 on breast cancer cells is studied, and the specific method is as follows:
human breast cancer cells (MCF-7) were selected, and the toxicity of MCF-7 (measured 24 hours after the addition of carbon dots to cells) was measured at concentrations of 0, 0.1, 0.2, 0.4, 0.6, 0.8, 1.0, 2.0, and 3.0mg/mL using MTT assay using a microplate reader, and the results are shown in FIG. 4 b. The experimental result shows that the survival rate of cells still exceeds 90% under the condition that the concentration of the carbon dots of the impatiens balsamina is far greater than the Minimum Inhibitory Concentration (MIC) of gram-positive bacteria, which indicates that the carbon dots of the impatiens balsamina have good biocompatibility.
3. The cytotoxicity of the obtained natural plant impatiens balsamina carbon dot prepared in example 1 on immortalized keratinocytes is studied by the following specific method:
human immortalized keratinocytes (HaCaT) were selected, and the toxicity of carbon dots to HaCaT (measured 24 hours after the addition of carbon dots to cells) was measured at concentrations of 0, 0.1, 0.2, 0.4, 0.6, 0.8, 1.0, 2.0, 3.0mg/mL by MTT assay using a microplate reader, and the results are shown in FIG. 4 c. The experimental result shows that the survival rate of cells still exceeds 90% under the condition that the concentration of the carbon dots of the impatiens balsamina is far greater than the Minimum Inhibitory Concentration (MIC) of gram-positive bacteria, which indicates that the carbon dots of the impatiens balsamina have good biocompatibility.
4. The cytotoxicity of the carbon dots of the obtained natural plant impatiens balsamina prepared in example 1 on macrophages is studied, and the specific method is as follows:
macrophages (RAW) were selected and toxicity of carbon spots to RAW (measured 24 hours after addition of carbon spots to cells) was measured at concentrations of 0, 0.1, 0.2, 0.4, 0.6, 0.8, 1.0, 2.0, 3.0mg/mL using MTT assay using a microplate reader, and the results are shown in the figure. The experimental result shows that the survival rate of the cells still exceeds 90% when the concentration of the carbon dots of the impatiens balsamina is higher than the Minimum Inhibitory Concentration (MIC) of gram-positive bacteria, which indicates that the carbon dots of the impatiens balsamina have good biocompatibility.
Fourthly, the carbon dots of the obtained natural plant impatiens balsamina prepared in the embodiment 1 of the invention have the effect of imaging gram-positive bacteria and gram-negative bacteria
1. Imaging effect of carbon point of natural plant impatiens balsamina on gram-positive bacteria
(1) The method for the imaging effect of the carbon points of the natural plant impatiens balsamina on staphylococcus aureus comprises the following steps:
staphylococcus aureus in the growth exponential phase was diluted to a turbidity of about 0.5 at 600nm, added to the carbon spot of impatiens balsamina of example 1 to a final concentration of 0.5mg/mL, cultured in an incubator at 37 ℃ for two hours, then washed 3 times with PBS solution, 10. mu.L was dropped onto a slide glass and observed under a fluorescence inverted microscope.
The results are shown in FIG. 5, which shows that the carbon dots of impatiens balsamina can be imaged on Staphylococcus aureus.
(2) The imaging effect of the carbon dots of the natural plant impatiens balsamina on the enterococcus faecium is realized by the following method:
enterococcus faecium in growth exponential phase is diluted to turbidity of about 0.5 at 600nm, carbon dots of impatiens balsamina in example 1 are added to final concentration of 0.5mg/mL, cultured in an incubator at 37 ℃ for two hours, then washed 3 times with PBS solution, 10. mu.L of the solution is dropped onto a slide glass and observed under a fluorescence inverted microscope.
The results are shown in FIG. 5, which indicates that the carbon dots of impatiens balsamina can be imaged on enterococcus faecium.
(3) The method for representing the effect of the carbon point of the natural plant impatiens balsamina on the bacillus subtilis comprises the following steps:
bacillus subtilis in growth exponential phase was diluted to a turbidity of about 0.5 at 600nm, added to the carbon spot of impatiens balsamina of example 1 to a final concentration of 0.5mg/mL, cultured in an incubator at 37 ℃ for two hours, then washed 3 times with PBS solution, 10. mu.L was dropped onto a slide glass and observed under a fluorescence inverted microscope.
The results are shown in FIG. 5, which shows that the carbon dots of impatiens balsamina can be visualized against Bacillus subtilis.
2. The carbon dot of natural plant impatiens balsamina has the effect of imaging gram-negative bacteria
(1) The method for expressing the effect of carbon points of natural plant impatiens balsamina on escherichia coli comprises the following steps:
coli in growth exponential phase was diluted to a turbidity of about 0.5 at 600nm, added to the carbon spot of impatiens balsamina of example 1 to a final concentration of 0.5mg/mL, incubated at 37 ℃ for two hours in an incubator, then washed 3 times with PBS solution, 10 μ L was dropped onto a slide glass and observed under a fluorescence inverted microscope.
The results are shown in FIG. 6, which shows that the carbon dots of impatiens balsamina cannot be visualized against E.coli.
(2) The method for the imaging effect of the carbon points of the natural plant impatiens balsamina on the salmonella comprises the following steps:
salmonella in the growth exponential phase was diluted to a turbidity of about 0.5 at 600nm, added to the carbon spot of impatiens balsamina of example 1 to a final concentration of 0.5mg/mL, cultured in an incubator at 37 ℃ for two hours, then washed 3 times with PBS solution, and 10. mu.L was dropped onto a slide glass and observed under a fluorescence inverted microscope.
The results are shown in FIG. 6, which shows that carbon spots of impatiens balsamina cannot be visualized for Salmonella.
(3) The method for the imaging effect of the carbon point of the natural plant impatiens balsamina on the pseudomonas aeruginosa comprises the following steps:
pseudomonas aeruginosa in the growth exponential phase was diluted to a turbidity of about 0.5 at 600nm, added to the carbon spot of impatiens balsamina of example 1 to a final concentration of 0.5mg/mL, incubated in an incubator at 37 ℃ for two hours, then washed 3 times with PBS solution, 10. mu.L was dropped onto a slide and observed under a fluorescence inverted microscope.
The results are shown in FIG. 6, which shows that the carbon dots of impatiens balsamina cannot be visualized against Pseudomonas aeruginosa.
While preferred embodiments of the present invention have been described, additional variations and modifications in those embodiments may occur to those skilled in the art once they learn of the basic inventive concepts. Therefore, it is intended that the appended claims be interpreted as including preferred embodiments and all such alterations and modifications as fall within the scope of the invention.
It will be apparent to those skilled in the art that various changes and modifications may be made in the present invention without departing from the spirit and scope of the invention. Thus, if such modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include such modifications and variations.
Claims (10)
1. A preparation method of natural plant impatiens balsamina carbon dots is characterized by comprising the following steps:
step 1, adding impatiens balsamina powder into a solvent, performing ultrasonic treatment, and reacting in a hydrothermal reaction kettle for 5-10h at the reaction temperature of 200-280 ℃;
the solvent is a mixed solution of a eutectic solvent and pure water;
step 2, after cooling to room temperature, centrifuging at 10000rpm of 8000-.
2. The method for preparing carbon dots of the natural plant impatiens balsamina as claimed in claim 1, wherein in step 1, the eutectic solvent consists of choline chloride and acetamide in a molar ratio of 1:1-2, or choline chloride and urea in a molar ratio of 1:1-2, or choline chloride and malonic acid in a molar ratio of 1:1-2, or choline chloride and acetic acid in a molar ratio of 1:1-2, or choline chloride and formic acid in a molar ratio of 1:1-2, or choline chloride and malic acid in a molar ratio of 1:1-2, or tetrabutylammonium bromide and glycerol in a molar ratio of 1-2:1-3, or tetrabutylammonium bromide and ethylene glycol in a molar ratio of 1:1-2, or tetrabutylammonium bromide and acetic acid in a molar ratio of 1:1-2, or the ammonium bromide tetrabutyl and the malonic acid according to the molar ratio of 1: 1-2.
3. The method for preparing carbon dots of the natural plant impatiens balsamina as claimed in claim 2, wherein the eutectic solvent consists of choline chloride and acetamide in a molar ratio of 1: 1.
4. The method for preparing natural plant impatiens carbon dots according to claim 1, wherein in step 1, the impatiens powder is obtained by drying and pulverizing impatiens petals and sieving.
5. The method for preparing carbon dots of impatiens balsamina as claimed in claim 1, wherein the ultrasonic treatment time in step 1 is 10-30 min.
6. The method for preparing natural plant impatiens balsamina carbon dots according to claim 1, wherein the mass ratio of the impatiens balsamina powder to the solvent in step 1 is 1: 10-30.
7. The method for preparing carbon dots of impatiens balsamina as claimed in claim 1, wherein the solvent contains pure water in an amount of 0-30% by volume.
8. The carbon dots of the natural plant impatiens balsamina prepared by the preparation method as set forth in any one of claims 1 to 7.
9. Use of the carbon dots of the natural plant impatiens balsamina as claimed in claim 8 for the identification of gram-positive bacteria.
10. Use of the natural plant impatiens balsamina carbon dots of claim 8 for antibacterial purposes.
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| CN114381262A (en) * | 2022-01-28 | 2022-04-22 | 河北科技大学 | Carbon quantum dot fluorescent probe and preparation method and application thereof |
| CN116022767A (en) * | 2023-02-15 | 2023-04-28 | 山东大学 | Carbon dots derived from herba Scutellariae Barbatae and herba Hedyotidis Diffusae, and preparation method and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114381262A (en) * | 2022-01-28 | 2022-04-22 | 河北科技大学 | Carbon quantum dot fluorescent probe and preparation method and application thereof |
| CN116022767A (en) * | 2023-02-15 | 2023-04-28 | 山东大学 | Carbon dots derived from herba Scutellariae Barbatae and herba Hedyotidis Diffusae, and preparation method and application thereof |
| CN116022767B (en) * | 2023-02-15 | 2024-05-24 | 山东大学 | Carbon dots derived from herba Scutellariae Barbatae and herba Hedyotidis Diffusae, and preparation method and application thereof |
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