CN106323931B - The method of detection using yeast bacterium as main carbon source microwave rapid synthesis carbon dots and for pH value of solution - Google Patents
The method of detection using yeast bacterium as main carbon source microwave rapid synthesis carbon dots and for pH value of solution Download PDFInfo
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Abstract
The invention belongs to chemical application technology fields, and in particular to it is a kind of using yeast bacterium as main carbon source microwave rapid synthesis carbon dots solution and be used for pH value of solution detection.The method that the present invention detects solution ph includes the following steps: that using yeast bacterium as main carbon source, carbon source supplemented by ethylenediamine, water is dispersion liquid, under microwave heating condition, synthesizes carbon dots solution.Obtain the carbon dots solution with fluorescent characteristic；The carbon dots of synthesis have fluorescent characteristic, and maximum excitation wavelength is 390 nm, and maximum emission wavelength is 460 nm；Carbon dots solution from after the solution reaction of different pH, it is in a linear relationship between the fluorescence intensity level of carbon dots solution and the pH of solution (I F =354.66-19.00 pH), the range of linearity of detection is 1.83-11.86.
Description
Technical field
The invention belongs to chemical application technology fields, and in particular to one kind is auxiliary by main carbon source ethylenediamine of yeast bacterium
Carbon source synthesizes carbon dots solution under microwave heating condition；Utilize the fluorescent value of carbon dots solution and solution after the solution reaction of different pH
Difference can detecte out the pH of solution.
Background technique
Fluorescent carbon point is most popular one of carbon nanomaterial after fullerene, carbon nanotube and graphene.This kind is received
Rice material overcomes certain disadvantages of traditional quantum dot, not only has excellent optical property and small size property, but also have
Good biocompatibility, it is easy to accomplish it is surface-functionalized, biochemical sensitive, imaging analysis, environment measuring, photocatalysis technology,
The fields such as metal ion detection and pharmaceutical carrier have good application potential.Synthetic method includes electrochemical process, and hydro-thermal method is micro-
Wave method, ultrasonic method etc..There is certain difficulty currently, realizing and being uniformly synthesized carbon dots still.It is more uniform that the present invention is directed to utilizations
Yeast bacterium is carbon source, to synthesize the better carbon dots of uniformity.
Early stage people utilize electrochemical measurement pH value, but due to the impedance of traditional glass electrode height, cracky and not
It is preferably used for the measurement of pH in fluorine-containing solution, and has " sodium error " in high alkalinity environment, encounters electrochemistry in pH value measurement
Many problems.Therefore, people continually develop new pH measurement method in recent years.It is fluorescence spy the present invention is based on newly synthesized carbon dots
Needle, the method for establishing detection pH value of solution.This method has easy to operate, the features such as detection range is wide.
Summary of the invention
The main object of the present invention is that the synthesis of existing carbon dots and the detection technique of pH value of solution and method are shown
The some defects come propose that a kind of using the uniform yeast bacterium of volume is main carbon source, equal in conjunction with the preparation of microwave method synthetic technology
The good carbon dots of even property and the detection for being used for pH value of solution.This method has many characteristics, such as that easy to operate, pH detection range is wide.
Purpose to realize the present invention, the technical solution adopted by the present invention are as follows:
A method of using yeast bacterium as main carbon source microwave rapid synthesis carbon dots and for the detection of pH value of solution, step
It is as follows:
(1) using yeast bacterium as main carbon source, ethylenediamine is auxiliary carbon source, first disperses second two for the yeast separated
Mixed solution is obtained in amine aqueous solution, then by mixed solution microwave heating treatment, with the water dissolution as thick solution of carbon dots, warp after reaction
Final carbon dots solution is obtained after the processing such as centrifugation, filtering；
(2) determine carbon dots solution fluorescence intensity level (I F ) and the pH value of solution between quantitative relationship；
(3) according to the relationship between the pH value of solution and the fluorescent value of carbon dots solution, it can detecte out the pH of solution to be measured
Value.
In step (2) carbon dots solution fluorescence intensity level (I F ) and the pH value of solution between quantitative relationship are as follows:I F =
354.66-19.00 pH (related coefficientr=0.9964)。
The concentration of the ethylenediamine is 3.0 mol/L.
It is middle high fire by the firepower of mixed solution microwave heating treatment described in step (1).
It is 25 min by the mixed solution microwave heating treatment time described in step (1).
Described in step (1), the thick solution of obtained carbon dots is respectively placed in centrifuge tube first and is centrifuged at a high speed, then
Obtained supernatant is carried out to be filtered under diminished pressure processing, finally obtains the transparent carbon dots solution of brown color.
Described in step (1), the centrifugal speed to the centrifugal treating of the thick solution of carbon dots is 10000 revs/min, when centrifugation
Between be 15 minutes.
The range of the detectable pH value of solution is 1.83-11.86.
The carbon dots average grain diameter of the synthesis is 3.8 nm.
The carbon dots of the synthesis have fluorescent characteristic, and maximum excitation wavelength is 390 nm, maximum emission wavelength 460
nm。
Detailed description of the invention
Fig. 1 is the fluorescence spectra of the carbon dots solution of preparation of the embodiment of the present invention.
Fig. 2 is the TEM figure of the carbon dots solution of preparation of the embodiment of the present invention.
Fig. 3 is grain size distribution in the TEM of the carbon dots solution of preparation of the embodiment of the present invention.
Fig. 4 is that the carbon dots solution of preparation of the embodiment of the present invention detects the figure of fluorescence intensity changes of different pH solution (1-11 is molten
The pH of liquid is respectively 1.83,2.91,3.71,4.70,5.72,6.80,7.87,8.81,9.84,10.81,11.86).
Specific embodiment:
Following embodiment is intended to further illustrate the present invention, rather than limitation of the invention further.
(1) culture and extraction of yeast bacterium
The ingredient of culture medium: 20.0 g of glucose, 1000 mL of distilled water.
The glucose for accurately weighing 20.0 g is added in the beaker containing 900 mL distilled water, is allowed to glass bar stirring
Sufficiently dissolution, is settled to 1000 mL.20 mL glucose solutions are measured into triangular flask, adds tampon, wrapped up with brown paper.It will be to
The glucose solution of sterilizing is put into autoclave, in 121 °C of 20 min of sterilizing.Sterilized glucose solution is placed in sterile work
Make in platform, opens 2 h of ultraviolet lamp sterilization.In strict accordance with sterility requirements, by Angel high activity dried yeast (the Angel ferment of 0.020 g
Female limited liability company) it is inoculated into glucose solution, 24 h of last 37 °C of cultures.
The saccharomycete for taking one bottle of above-mentioned culture is sub-packed in the centrifuge tube of 45 mL, with 10000 revs/min of speed from
The heart 10 minutes；Precipitating is taken after centrifugation, saccharomycete is deposited in centrifugation bottom of the tube in solid form at this time；By each centrifuge tube
In saccharomycete mixed respectively with the ethylenediamine solution of 1.0 mL, 3.0 mol/L.
(2) preparation of carbon dots
12 above-mentioned centrifuge tubes are taken, solution therein are transferred in the conical flask of 250 mL, then are added 8 toward conical flask
The ethylenediamine solution of mL, 3.0 mol/L, until final volume is 20 mL.Conical flask is transferred in micro-wave oven, middle high fire, microwave
Heat 25 min.Conical flask is taken out after reaction, is dissolved with 10 mL water, and acquired solution is the thick solution of carbon dots.Carbon dots are slightly molten
Liquid is centrifuged 15 minutes first with 10000 revs/min, takes supernatant, then is obtained by filtration with the filter membrane vacuum decompression in 0.2 μm of aperture
Carbon dots solution.As shown in Figure 1, maximum excitation wavelength is 390 nm, maximum emission wavelength is 460 nm.And to the pattern etc. of carbon dots
TEM characterization has been carried out, as shown in Fig. 2, the carbon dots good dispersion of synthesis, and more uniformly.Particle diameter distribution result (such as Fig. 3) shows
3.82 nm of carbon dots size is in left and right.
(3) the preparation condition optimization of carbon dots
The influence of concentration, volume, microwave heating time and the firepower of ethylenediamine to synthesis carbon dots is inquired into, mainly from pairing
At carbon dots solution fluorescence intensity influence investigated.
The concentration of ethylenediamine has an impact to the fluorescence intensity of carbon dots solution, and the concentration for having inquired into ethylenediamine is respectively 1.0
To the fluorescence intensity of the carbon dots solution of synthesis when mol/L, 3.0 mol/L, 5.0 mol/L, 8.0 mol/L, 10.0 mol/L
It influences.When the concentration of ethylenediamine is 3.0 mol/L, 5.0 mol/L, the fluorescence of carbon dots solution is stronger, and when ethylenediamine concentration is
When 3.0 mol/L, the fluorescence of carbon dots solution is most strong, and final preferred concentration is 3.0 mol/L.
The volume of ethylenediamine has an impact to the fluorescence intensity of carbon dots solution, inquired into ethylenediamine volume be respectively 10 mL,
The influence of 15 mL, 20 mL, 25 mL, 30 mL, 35 mL to the fluorescence intensity of the carbon dots solution of synthesis.When the volume of ethylenediamine
The fluorescence of carbon dots solution is stronger when for 20 mL, 25 mL, and when the volume of ethylenediamine is 20 mL, the fluorescence of carbon dots solution is most
By force, final preferred volume is 20 mL.
The quantity of bacterium has an impact to the fluorescence intensity of carbon dots solution.The amount of bacterium is indicated with the volume of inoculum
Influence to the fluorescence intensity of carbon dots solution.Probed into culture solution volume be respectively 20 mL, 40 mL, 60 mL, 80 mL,
Influence of 100 mL to the fluorescence intensity of the carbon dots solution of synthesis.When the volume of culture solution is 60 mL, 80 mL, carbon dots solution
Fluorescence it is stronger, and when the volume of culture solution is 60 mL, the fluorescence of carbon dots solution is most strong, the volume of final preferably culture solution
For 60 mL.
Microwave firepower has an impact to the fluorescence intensity of carbon dots solution.Probed into microwave firepower be respectively in low fire, moderate heat, in
To the influence of the fluorescence intensity of synthesis carbon dots solution when Gao Huo, high fire.When microwave firepower is moderate heat, middle high fire, carbon dots solution
Fluorescence it is stronger, and when microwave firepower is middle high fire, the fluorescence of carbon dots solution is most strong, and final preferably microwave firepower is middle height
Fire.
Microwave heating time has an impact to the fluorescence intensity of carbon dots solution.Having probed into microwave heating time is respectively 10
To the influence of the fluorescence intensity of synthesis carbon dots solution when min, 15min, 20 min, 25 min, 30 min.Work as microwave heating time
When for 20 min, 25 min, the fluorescence of carbon dots solution is stronger, and when microwave heating time is 25min, the fluorescence of carbon dots solution
Most strong, the final preferably microwave time is 25 min.
(4) detection parameters of the carbon dots to pH value of solution
The centrifuge tube of 11 2.0 mL is taken, A, B, C, D, E, F, G, H, I, J, No. K are put on respectively, into A-K branch centrifuge tube
The carbon dots solution that 1.6 mL diluted 50 times in advance is added, then sequentially adds 400 μ L difference pH's into A-K centrifuge tube
Britton-Robinson buffer solution, pH value is respectively 1.83,2.91,3.71,4.70,5.72,6.80,7.87,8.81,
9.84,10.81,11.86.After reacting 30 min, the solution in each centrifuge tube is successively surveyed in sepectrophotofluorometer
Examination records fluorescence spectrum value of carbon dots in the presence of different BR buffer solutions, as a result sees Fig. 4, fluorescence intensity level (I F ) and solution
PH value between quantitative relationship are as follows:I F =354.66-19.00 pH (r=0.9964)。
It should also be noted that, specific embodiments of the present invention are used only to exemplary illustration, do not limit in any way
Determining protection scope of the present invention, the related technical personnel of this field can be improved or be changed according to above-mentioned some explanations, but
All these improvements and changes all should belong to the protection scope of the claims in the present invention.
Claims (7)
1. a kind of method of the detection using yeast bacterium as main carbon source microwave rapid synthesis carbon dots and for pH value of solution, feature exist
In following steps:
(1) using yeast bacterium as main carbon source, ethylenediamine is auxiliary carbon source, and it is molten first to disperse ethylenediamine for the yeast separated
Mixed solution being obtained in liquid, then by 25 min of mixed solution microwave heating treatment, being dissolved after reaction with water is the thick solution of carbon dots,
Final carbon dots solution is obtained after centrifugation, filtration treatment；
(2) determine carbon dots solution fluorescence intensity level (I F ) and the pH value of solution between quantitative relationship；The fluorescence of carbon dots solution
Intensity value (I F ) and the pH value of solution between quantitative relationship are as follows:I F =354.66-19.00 pH；
(3) according to the relationship between the pH value of solution and the fluorescent value of carbon dots solution, the pH value that can detecte out solution to be measured is
1.83-2.91。
2. it is according to claim 1 using yeast bacterium as main carbon source microwave rapid synthesis carbon dots and be used for pH value of solution detection
Method, it is characterised in that: the concentration of ethylenediamine be 3.0 mol/L.
3. it is according to claim 1 using yeast bacterium as main carbon source microwave rapid synthesis carbon dots and be used for pH value of solution detection
Method, it is characterised in that: step (1) firepower by mixed solution microwave heating treatment is middle high fire.
4. it is according to claim 1 using yeast bacterium as main carbon source microwave rapid synthesis carbon dots and be used for pH value of solution detection
Method, it is characterised in that: in step (1), first by the thick solution of obtained carbon dots be respectively placed in centrifuge tube carry out high speed centrifugation
Separation, then obtained supernatant is carried out to be filtered under diminished pressure processing, finally obtain the transparent carbon dots solution of brown color.
5. it is according to claim 1 using yeast bacterium as main carbon source microwave rapid synthesis carbon dots and be used for pH value of solution detection
Method, it is characterised in that: in step (1) to the centrifugal speed of the centrifugal treating of the thick solution of carbon dots be 10000 revs/min, from
The heart time is 15 minutes.
6. it is according to claim 1 using yeast bacterium as main carbon source microwave rapid synthesis carbon dots and be used for pH value of solution detection
Method, it is characterised in that: the carbon dots average grain diameter of synthesis be 3.8 nm.
7. it is according to claim 1 using yeast bacterium as main carbon source microwave rapid synthesis carbon dots and be used for pH value of solution detection
Method, it is characterised in that: the carbon dots of synthesis have fluorescent characteristic, and maximum excitation wavelength is 390 nm, and maximum emission wavelength is
460 nm。
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| CN107916105B (en) * | 2017-11-17 | 2020-11-10 | 山西大学 | Red fluorescent carbon quantum dot for detecting intracellular pH and preparation method thereof |
| CN108083257B (en) * | 2017-12-28 | 2021-07-30 | 湖南科技大学 | Method for preparing fluorescent carbon dots by taking chloroplasts as carbon sources |
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| CN109324028B (en) * | 2018-11-19 | 2021-04-09 | 湖南科技大学 | Method for detecting Cr (VI) by using ethylenediamine and nitric acid as raw materials to rapidly synthesize carbon dot solution through microwave |
| CN110172344A (en) * | 2019-04-28 | 2019-08-27 | 山西大学 | For monitoring the fluorescent carbon quantum dot and its preparation method and application of acidic environment pH |
| CN111999275B (en) * | 2020-08-25 | 2022-06-17 | 中南民族大学 | Method for rapidly and quantitatively measuring pH value and/or uric acid |
| CN112113944A (en) * | 2020-09-30 | 2020-12-22 | 江南大学 | Preparation of emission spectrum variable carbon dots and application thereof in detection of pH value |
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