CN108083257B - Method for preparing fluorescent carbon dots by taking chloroplasts as carbon sources - Google Patents
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- CN108083257B CN108083257B CN201711458689.2A CN201711458689A CN108083257B CN 108083257 B CN108083257 B CN 108083257B CN 201711458689 A CN201711458689 A CN 201711458689A CN 108083257 B CN108083257 B CN 108083257B
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- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
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Abstract
The invention belongs to the technical field of chemical application, and particularly relates to a hydrothermal method for synthesizing a fluorescent carbon dot solution by taking chloroplast as a carbon source and for detecting pH. The method for detecting the pH value of the solution comprises the following steps: synthesizing a fluorescent carbon dot solution by taking chloroplast as a carbon source and water as a dispersion under the heating condition of a hydrothermal reaction kettle. Obtaining a carbon dot solution with fluorescence characteristics; after the fluorescent carbon dot solution reacts with solutions with different pH values, the fluorescence intensity value and the pH value of the fluorescent carbon dot solution show good linear relation IF395.44-10.36x, wherein IFThe method has the advantages of simple process, rapid preparation of the fluorescent carbon dots and accurate determination of the pH value of the solution, and can detect the pH value of the solution within the range of 1.83-10.38.
Description
Technical Field
The invention belongs to the technical field of chemical application, and particularly relates to a method for synthesizing a fluorescent carbon dot solution by using chloroplast as a carbon source and a hydrothermal method; the pH value of the solution can be detected by utilizing the difference of the fluorescence intensity values of the reaction of the fluorescent carbon dot solution and the solutions with different pH values.
Background
The fluorescent carbon dot is a novel nano material with the diameter less than 10nm, and becomes a research hotspot of a carbon nano family due to the characteristics of low toxicity, stable chemical property, excellent water solubility, photobleaching resistance, easy functionalization and the like. The excellent performances of the fluorescent carbon dots enable the fluorescent carbon dots to have wide application prospects in the fields of biological markers, environmental analysis, photoelectric sensing and the like. At present, the realization of uniform carbon point synthesis is still difficult, and the invention aims to utilize chloroplast contained in most plant leaves as a carbon source to synthesize carbon points with better uniformity.
Early people utilized electrochemical methods to measure pH, but because the impedance of the traditional glass electrode is high, the traditional glass electrode is easy to break and is not suitable for measuring pH in fluorine-containing solution, and the traditional glass electrode has sodium error in high alkaline environment, the electrochemistry has many problems in pH measurement. Therefore, new pH measurement methods have been developed in recent years. The invention establishes a method for detecting the pH value of a solution based on a newly synthesized fluorescent carbon dot as a fluorescent probe. The method has the characteristics of simple operation, wide detection range and the like.
The present invention has been made in view of this situation.
Disclosure of Invention
The technical problem to be solved by the invention is to overcome the defects of the prior art, provide a method for preparing fluorescent carbon dots by taking chloroplast as a carbon source and application thereof,
in order to solve the technical problems, the invention adopts the technical scheme that:
the invention mainly aims to overcome the defects of the existing fluorescent carbon dot synthesis and solution pH detection technology and method, and provides a method for synthesizing and preparing a fluorescent carbon dot with good uniformity by combining a hydrothermal method and taking chloroplast in most plants as a carbon source and using the chloroplast in the plants as a carbon source for pH detection. The method has the characteristics of simple operation, wide pH detection range and the like.
A method for preparing fluorescent carbon dots by taking chloroplasts as carbon sources comprises the following steps:
s1: extracting chloroplast from fresh plant leaves serving as a raw material, grinding, filtering and centrifuging to extract chloroplast suspension;
s2: taking the chloroplast suspension obtained in S1 as a carbon source, heating the chloroplast suspension in a hydrothermal reaction kettle, synthesizing a primary fluorescent carbon dot solution, and performing centrifugal separation treatment to obtain a fluorescent carbon dot solution;
s3: detecting the fluorescence intensity value of the fluorescent carbon dot solution obtained in the step S2 after the fluorescent carbon dot solution reacts with the solutions with different pH values;
s4: listing the corresponding relation between the fluorescence intensity value and the pH value according to the fluorescence intensity value of the fluorescent carbon dot solution after the reaction with the solutions with different pH values;
s5: and determining the pH value of the unknown solution according to the corresponding relation between the pH value and the fluorescence intensity value.
In the invention, the plant leaves in S1 are green vegetable leaves, and the green vegetable leaves are spinach leaves; s1, grinding, filtering and centrifuging to extract chloroplast suspension, wherein the chloroplast suspension comprises the following steps: adding isotonic solution into fresh spinach leaves as a raw material, grinding, filtering, and removing precipitates to obtain ground homogenate; then, centrifuging the homogenate to remove the lower-layer precipitate and taking the supernatant; and centrifuging the supernatant, taking the supernatant precipitate, and adding an isotonic solution into the supernatant precipitate to obtain a chloroplast suspension, wherein the isotonic solution is a sodium chloride solution, the molar concentration of the sodium chloride solution is 0.35mol/L, and the volume of the chloroplast suspension is 10 mL.
In the invention, the step of heating the hydrothermal reaction kettle in S2 is as follows: transferring the obtained chloroplast suspension into a hydrothermal reaction kettle, and heating the reaction kettle in a drying oven to obtain a rough fluorescent carbon dot solution; and (3) heating the reaction kettle in an oven at the temperature of 150-250 ℃ for 1-3 h.
In the present invention, the centrifugation step in S2 is: and respectively placing the obtained fluorescent carbon dot solution into a centrifuge tube for centrifugal separation to obtain brown yellow transparent liquid, namely the fluorescent carbon dot solution, wherein the centrifuge tube has the centrifugal speed of 10000r/min and the centrifugal time of 20min when carrying out centrifugal separation.
In the invention, the fluorescence intensity value and the pH value in S4 show good linear relation IF395.44-10.36x, wherein IFFluorescence intensity values, x is the pH value.
An application of preparing a fluorescent carbon dot by taking chloroplast as a carbon source, a fluorescent carbon dot solution prepared by taking chloroplast as a carbon source is used for detecting the pH of the solution,
the application of the fluorescent carbon dot solution prepared by using the chloroplast as the carbon source is used for detecting the pH of the solution, and the pH range of the detectable solution is 1.83-10.38.
The synthesized fluorescent carbon dots have the fluorescent characteristic, the maximum excitation wavelength is 320nm, and the maximum emission wavelength is 439 nm.
The application of chloroplast as a carbon source in preparing a fluorescent carbon dot is used for detecting the pH of a solution and comprises the following steps:
heating chloroplast serving as a carbon source in an oven under a hydrothermal condition to synthesize a fluorescent carbon dot solution, and treating to obtain the fluorescent carbon dot solution; after the fluorescent carbon dot solution reacts with solutions with different pH values, detecting the fluorescence intensity value of the fluorescent carbon dot solution; and detecting the pH value of the solution according to the relation between the pH value of the solution and the fluorescence intensity value of the fluorescent carbon dot solution.
After the technical scheme is adopted, compared with the prior art, the invention has the following beneficial effects.
The method has simple process, can quickly prepare the fluorescent carbon dots, and can accurately measure the pH of the solution.
The following describes embodiments of the present invention in further detail with reference to the accompanying drawings.
Drawings
The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this application, illustrate embodiment(s) of the invention and together with the description serve to explain the invention without limiting the invention to its proper form. It is obvious that the drawings in the following description are only some embodiments, and that for a person skilled in the art, other drawings can be derived from them without inventive effort. In the drawings:
FIG. 1 is a fluorescence spectrum of a fluorescent carbon dot solution prepared according to an example of the present invention;
FIG. 2 is a transmission electron microscope image of a fluorescent carbon dot solution prepared according to an embodiment of the present invention;
FIG. 3 is a graph showing the change of fluorescence intensity of pH solutions for detecting fluorescent carbon dot solutions prepared by practicing the present invention;
FIG. 4 is a fluorescence line graph of a fluorescent carbon dot solution prepared according to an embodiment of the present invention.
It should be noted that the drawings and the description are not intended to limit the scope of the inventive concept in any way, but to illustrate it by a person skilled in the art with reference to specific embodiments.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments will be clearly and completely described below with reference to the embodiments of the present invention, and the following embodiments are used for illustrating the present invention and are not intended to limit the scope of the present invention.
Example 1
Extracting chloroplasts: taking spinach as an example: selecting fresh spinach leaves, washing with water for 2-3 times, and wiping to remove large veins. 10g of spinach leaves are accurately weighed, cut into pieces and put into a mortar, and 20mL of 0.35mol/L NaCl solution is added for grinding. The homogenate after grinding was filtered through six layers of gauze into a 100mL beaker and the filter residue was discarded. Transferring the filtered filtrate into a 5mL centrifuge tube, centrifuging for 10min at the rotating speed of 1000r/min, and discarding the precipitate. Transferring the supernatant into a 5mL centrifuge tube, continuously centrifuging for 20min at 3000r/min, removing the supernatant, taking the lower-layer precipitate, washing with 0.35mol/L NaCl solution for 2-3 times, and adding 10mL of 0.35mol/L NaCl solution to suspend the precipitate. A chloroplast-containing suspension was obtained.
Preparation of fluorescent carbon dots: the chloroplast suspension was transferred to a 10mL reaction vessel and placed in an oven at 200 ℃ for two hours. Finally, 8mL of the primary fluorescent carbon dot solution was obtained. Centrifuging the primary fluorescent carbon dot solution for 20min at 10000r/min, and taking supernatant, namely the fluorescent carbon dot solution. As shown in FIG. 1, the maximum excitation wavelength is 320nm and the maximum emission wavelength is 439 nm. And performing TEM representation on the appearance of the fluorescent carbon dots. As shown in FIG. 2, the synthesized fluorescent carbon dots have good dispersibility and are relatively uniform.
Detection parameters of the fluorescent carbon dots on the pH of the solution: 10 5.0mL centrifuge tubes were labeled A, B, C, D, E, F, G, H, I, J, 1.60mL of 6-fold pre-diluted fluorescent carbon spot solution was added to the A-J centrifuge tubes, and 400uL of Britton-Robinson buffer solutions of different pH values, 1.81, 2.87, 3.78, 4.78, 5.72, 6.80, 7.96, 8.95, 9.91, and 10.38, respectively, were added to the A-J centrifuge tubes. After reacting for 30min, sequentially testing the solutions in the centrifuge tubes in a fluorescence spectrophotometer, recording the fluorescence spectrum values of the fluorescent carbon points in the presence of different BR buffer solutions, and referring to FIG. 3, which is a fluorescence spectrum diagram of the fluorescent carbon points at different pH values, wherein a good linear relationship I is presented between the fluorescence intensity value and the pH valueF395.44-10.36x, the result is shown in fig. 4, and R is 0.9970.
Although the present invention has been described with reference to a preferred embodiment, it should be understood that various changes, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined by the appended claims.
Claims (3)
1. The application of the chloroplast as the carbon source to prepare the fluorescent carbon dots is characterized in that the chloroplast is used as the carbon source, a fluorescent carbon dot solution is synthesized by heating in an oven under a hydrothermal condition, and the fluorescent carbon dot solution is obtained after treatment; after the fluorescent carbon dot solution reacts with solutions with different pH values, detecting the fluorescence intensity value of the fluorescent carbon dot solution; according to the relation between the pH value of the solution and the fluorescence intensity value of the fluorescent carbon dot solution, the pH value of the solution can be detected, the synthesized fluorescent carbon dot has the fluorescence characteristic, the maximum excitation wavelength is 320nm, the maximum emission wavelength is 439nm, the range of the detectable solution pH is 1.83-10.38, and the specific process is as follows:
s1: extracting chloroplast from fresh plant leaves serving as a raw material, grinding, filtering and centrifuging to extract chloroplast suspension;
s2: taking the chloroplast suspension obtained in S1 as a carbon source, transferring the chloroplast suspension into a hydrothermal reaction kettle, heating the hydrothermal reaction kettle in an oven at the temperature of 150-250 ℃ for 1-3 h to synthesize a primary fluorescent carbon dot solution, and performing centrifugal separation to obtain a fluorescent carbon dot solution;
s3: detecting the fluorescence intensity value of the fluorescent carbon dot solution obtained in the step S2 after the fluorescent carbon dot solution reacts with the solutions with different pH values;
s4: listing the corresponding relation between the fluorescence intensity value and the pH value according to the fluorescence intensity value of the fluorescent carbon dot solution after the reaction with the solutions with different pH values, wherein the fluorescence intensity value and the pH value have good linear relation IF (intermediate frequency) of 395.44-10.36x, wherein IF is the fluorescence intensity value, and x is the pH value;
s5: and determining the pH value of the unknown solution according to the corresponding relation between the pH value and the fluorescence intensity value.
2. The use of claim 1, wherein the plant leaves in S1 are green vegetable leaves, and the green vegetable leaves are spinach leaves; s1, grinding, filtering and centrifuging to extract chloroplast suspension, wherein the chloroplast suspension comprises the following steps: adding isotonic solution into fresh spinach leaves as a raw material, grinding, filtering, and removing precipitates to obtain ground homogenate; then centrifuging the homogenate to remove the lower-layer precipitate and taking the supernatant; and centrifuging the supernatant, taking the supernatant precipitate, and adding an isotonic solution into the supernatant precipitate to obtain a chloroplast suspension, wherein the isotonic solution is a sodium chloride solution, the molar concentration of the sodium chloride solution is 0.35mol/L, and the volume of the chloroplast suspension is 10 mL.
3. The use of claim 1, wherein the step of centrifuging in S2 comprises: and respectively placing the obtained fluorescent carbon dot solution into a centrifuge tube for centrifugal separation to obtain brown yellow transparent liquid, namely the fluorescent carbon dot solution, wherein the centrifuge tube has the centrifugal speed of 10000r/min and the centrifugal time of 20min when carrying out centrifugal separation.
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| CN109324027B (en) * | 2018-11-18 | 2021-04-09 | 湖南科技大学 | Method for preparing fluorescent carbon dot detection aureomycin by taking p-phenylenediamine and acetic acid as carbon sources |
| CN109880620A (en) * | 2019-03-02 | 2019-06-14 | 福建医科大学 | It is a kind of using biomass as the preparation and application of the green fluorescence carbon quantum dot of presoma |
| CN109777412B (en) * | 2019-03-21 | 2021-12-31 | 山西大学 | Double-emission fluorescent carbon dot and preparation method and application thereof |
| CN110117492B (en) * | 2019-04-17 | 2021-12-10 | 长春工业大学 | Fluorescent carbon dot and preparation method and application thereof |
| CN111977632A (en) * | 2019-05-22 | 2020-11-24 | 东北林业大学 | Green preparation method of spinach-based multicolor luminescent carbon quantum dots |
| CN110396406B (en) * | 2019-06-11 | 2021-10-19 | 华南农业大学 | Chlorophyll carbon dot, preparation method thereof and application thereof in time detection label |
| CN110373185B (en) * | 2019-08-02 | 2022-04-12 | 河南师范大学 | Selective detection of Bi by synthesizing fluorescent carbon dots by taking broussonetia papyrifera leaves as carbon source3+Method (2) |
| CN110308126A (en) * | 2019-08-02 | 2019-10-08 | 河南师范大学 | A method of using chinaberry hinge at fluorescence nano carbon dots cell imaging |
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