CN110044861A - The detection method of concentration of hydrogen peroxide - Google Patents
The detection method of concentration of hydrogen peroxide Download PDFInfo
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- CN110044861A CN110044861A CN201910384908.XA CN201910384908A CN110044861A CN 110044861 A CN110044861 A CN 110044861A CN 201910384908 A CN201910384908 A CN 201910384908A CN 110044861 A CN110044861 A CN 110044861A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- 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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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
- G01N2021/6432—Quenching
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
- G01N2021/6439—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" with indicators, stains, dyes, tags, labels, marks
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Abstract
The invention discloses the detection methods of concentration of hydrogen peroxide, comprising: Step 1: preparing the Hydrogen peroxide standard solution of carbon quantum dot solution, horseradish peroxidase solution, o-phenylenediamine solution and various concentration；Step 2: successively adding horseradish peroxidase solution, o-phenylenediamine solution and carbon quantum dot solution into Hydrogen peroxide standard solution respectively, after hybrid reaction, standard sample to be tested system is obtained；Step 3: examination criteria sample to be tested ties up to the fluorescence intensity at 442nm and 573nm, and establish fluorescence intensity I573/I442Linear relation between concentration of hydrogen peroxide；Step 4: preparing hydrogen peroxide solution to be measured, after its method hybrid reaction according to step 2, fluorescence intensity of the reactant at 442nm and 573nm is detected, according to linear relation, calculates the concentration of hydrogen peroxide in hydrogen peroxide solution to be measured.The present invention have many advantages, such as it is easy to operate, detection quickly and high sensitivity.
Description
Technical field
The present invention relates to hydrogen peroxide detection technique fields.It is more particularly related to a kind of concentration of hydrogen peroxide
Detection method.
Background technique
Hydrogen peroxide is a kind of inorganic Green Fine Chemical product, and hydrogen peroxide is widely used in weaving, papermaking, change
The fields such as work, electronics, environmental protection, food, cosmetics, medicine, mining and metallurgy and military project.However, being exposed to the peroxide of high concentration for a long time
Change in hydrogen environment, the product that long-term consumption remains the food of hydrogen peroxide or uses content of hydrogen peroxide exceeded, will seriously damage
Evil human health.Currently, mainly having chemoluminescence method, fluorescence method, spectrophotometry to the detection method of hydrogen peroxide both at home and abroad
And electrochemical method etc., but most methods there are operating methods it is complicated, the reaction time is long, sensitivity is not high the defects of.
Summary of the invention
It is an object of the present invention to provide a kind of detection methods of concentration of hydrogen peroxide, using carbon quantum dot as glimmering
Light probe detects the concentration of hydrogen peroxide in solution, and this method is easy to operate, detection is quick and high sensitivity, can be carried out in sample
The highly sensitive identification of hydrogen peroxide.
In order to realize purposes and other advantages according to the present invention, a kind of detection method of concentration of hydrogen peroxide is provided,
Include:
Step 1: preparing the mistake of carbon quantum dot solution, horseradish peroxidase solution, o-phenylenediamine solution and various concentration
Hydrogen oxide standard solution；
Step 2: successively adding isometric horseradish into the Hydrogen peroxide standard solution of same volume various concentration respectively
Peroxidase Solution, o-phenylenediamine solution and carbon quantum dot solution after hybrid reaction, obtain standard sample to be tested system；
Step 3: detecting the standard sample to be tested ties up to fluorescence intensity at 442nm and 573nm, and it is strong to establish fluorescence
Spend I573/I442Linear relation between concentration of hydrogen peroxide；
It is waited for Step 4: preparing the hydrogen peroxide unknown with the volume same concentrations of the Hydrogen peroxide standard solution in step 2
Solution is surveyed, after its method hybrid reaction according to step 2, detects fluorescence intensity of the reactant at 442nm and 573nm, root
According to fluorescence intensity I573/I442Linear relation between concentration of hydrogen peroxide calculates the mistake in hydrogen peroxide solution to be measured
Hydrogen peroxide concentration.
Preferably, the detection method of the concentration of hydrogen peroxide in step 1, prepares carbon quantum dot solution, horseradish
When the Hydrogen peroxide standard solution of Peroxidase Solution, o-phenylenediamine solution and various concentration, the solvent of use is Tris-
HCl buffer solution, pH 7.4, concentration 0.05mol/L.
Preferably, the detection method of the concentration of hydrogen peroxide, in step 1, the Hydrogen peroxide standard of various concentration
The preparation method of solution are as follows: the hydrogen peroxide stoste for being first 10mol/L with pure hydrogen peroxide compound concentration, by stoste Tris-
HCl buffer solution carries out gradient dilution to get the concentration of the Hydrogen peroxide standard solution of the various concentration is followed successively by 0mol/
L、7.5×10-5mol/L、1.5×10-4mol/L、2.25×10-4mol/L、3×10-4mol/L。
Preferably, the detection method of the concentration of hydrogen peroxide, in step 1, the concentration of carbon quantum dot solution is
5.3×10-6G/mL, the concentration of horseradish peroxidase solution are 1 × 10-5G/mL, the concentration of o-phenylenediamine solution are 3 × 10- 2mol/L。
Preferably, the detection method of the concentration of hydrogen peroxide, in step 1, carbon quantum dot the preparation method comprises the following steps:
The pancreatin of 2 parts by weight is dissolved in the ultrapure water of 10 parts by weight, after dispersion, is put it into reaction kettle, reacted at 180 DEG C
After 10h, take out, freeze-drying to get.
Preferably, the detection method of the concentration of hydrogen peroxide, in step 2 specifically: first respectively to same volume
Isometric horseradish peroxidase solution and o-phenylenediamine solution are added in the Hydrogen peroxide standard solution of various concentration, in 37
At DEG C after hybrid reaction 30min, then it is separately added into isometric carbon quantum dot solution, and be stirred with microsyringe, often
The lower hybrid reaction 1min of temperature.
Preferably, the detection method of the concentration of hydrogen peroxide, in step 3, the condition of fluorescence detection are as follows: excitation
Wavelength is 380nm, and excitation and transmite slit are 5nm.
Preferably, pancreatin is dissolved in ultrapure water when prepared by carbon quantum dot by the detection method of the concentration of hydrogen peroxide
In, it after dispersion, first puts it into magnetizer, the magnetization treatment 15min under the magnetic field strength of 12T is subsequently placed at 37 DEG C and protects
Warm 30min, then put it into super-pressure reactor tank, the ultra high pressure treatment 5min under 240MPa finally puts it into reaction again
In kettle, after reacting 10h at 180 DEG C, take out, freeze-drying.
The present invention is include at least the following beneficial effects:
The first, the present invention generates the hydroxyl with strong oxidizing property after interacting using horseradish peroxidase and hydrogen peroxide
O-phenylenediamine is oxidized to 2,3- diaminophenazine by base free radical, and 2,3- diaminophenazines can be quenched by self-filtering effect
The fluorescence of carbon quantum dot, and 2,3- diaminophenazines itself also have the fluorescence of some strength, it is possible thereby to pass through the glimmering of the two
Intensity ratio constructs the content of Ratio-type detection architecture detection hydrogen peroxide, and detection process is simple and convenient, high sensitivity, detection
Limit low, it can be achieved that mixing the rapid sensitive detection of hydrogen peroxide in sample, detection of the method disclosed by the invention to hydrogen peroxide
Limit can reach 3 × 10-5mol/L。
The second, using the carbon quantum dot of pancreatin synthesis as probe in detecting hydrogen peroxide high specificity, by other metal ions
Interfere less, testing result is accurate and reliable, and carbon quantum dot fluorescence property is excellent, and biocompatibility is preferable, facilitates detection, especially makes
With the exciting light of 380nm, preferable fluorescence spectrum effect is obtained.
Third, by pancreatin dispersion liquid be put into reaction kettle carry out hydrothermal synthesis reaction before, first by pancreatin dispersion liquid according to
It is secondary pancreatin to be made sufficiently to activate through magnetization, 37 DEG C of heat preservations and ultra high pressure treatment, magnetization treatment, improve the activity of pancreatin, 37 DEG C of guarantors
Temperature processing, can make sufficiently to react between each component of pancreatin, degrade, the solubility of pancreatin in water can be improved in ultra high pressure treatment, makes
The yield of carbon quantum dot is improved by 43.5% to 53.6%, and the detection limit of hydrogen peroxide is further decreased, up to 1.52 ×
10-5mol/L。
Further advantage, target and feature of the invention will be partially reflected by the following instructions, and part will also be by this
The research and practice of invention and be understood by the person skilled in the art.
Detailed description of the invention
Fig. 1 according to an embodiment of the present invention in standard sample to be tested system, excitation wavelength be 380nm when obtain it is glimmering
Light spectrogram；
Fig. 2 be according to an embodiment of the present invention in using concentration of hydrogen peroxide as abscissa, I573/I442For ordinate, draw
The standard curve of system.
Specific embodiment
Below with reference to embodiment and attached drawing, the present invention is described in further detail, to enable those skilled in the art's reference
Specification word can be implemented accordingly.
It should be noted that experimental method described in following embodiments is unless otherwise specified conventional method, institute
Reagent and material are stated, unless otherwise specified, is commercially obtained.
Embodiment 1:
The present invention provides a kind of detection method of concentration of hydrogen peroxide, comprising:
Step 1: being 7.4 with pH, concentration is the Tris-HCl buffer preparation carbon quantum dot solution, peppery of 0.05mol/L
The Hydrogen peroxide standard solution of root Peroxidase Solution, o-phenylenediamine solution and various concentration, wherein carbon quantum dot solution
Concentration is 5.3 × 10-6G/mL, the concentration of horseradish peroxidase solution are 1 × 10-5The concentration of g/mL, o-phenylenediamine solution is
3×10-2mol/L；The preparation method of the Hydrogen peroxide standard solution of various concentration are as follows: first prepared with pure hydrogen peroxide and ultrapure water
Concentration is the hydrogen peroxide stoste of 10mol/L, and stoste is carried out gradient dilution with the Tris-HCl buffer to get described
The concentration of the Hydrogen peroxide standard solution of various concentration is followed successively by 0mol/L, 7.5 × 10-5mol/L、1.5×10-4mol/L、
2.25×10-4mol/L、3×10-4mol/L；Carbon quantum dot the preparation method comprises the following steps: the pancreatin of 4g to be dissolved in the ultrapure water of 20ml
In, after dispersion, put it into reaction kettle, at 180 DEG C react 10h after, take out, freeze-drying to get.
Step 2: the horseradish of 100 μ l is successively added into the Hydrogen peroxide standard solution of the 100 above-mentioned various concentrations of μ l respectively
Peroxidase Solution, 100 μ l o-phenylenediamine solutions and 100 μ l carbon quantum dot solution after hybrid reaction, obtain standard sample to be tested
System；
Step 3: standard sample to be tested system is sequentially placed into RF-6000 sepectrophotofluorometer, excitation wave is set
A length of 380nm, excitation and transmite slit be 5nm, detect standard sample to be tested system tie up to it is glimmering at 442nm and 573nm
Luminous intensity, and establish fluorescence intensity I573/I442Linear relation between concentration of hydrogen peroxide；
It is waited for Step 4: preparing the hydrogen peroxide unknown with the volume same concentrations of the Hydrogen peroxide standard solution in step 2
Solution is surveyed, after its method hybrid reaction according to step 2, detects fluorescence intensity of the reactant at 442nm and 573nm, root
According to fluorescence intensity I573/I442Linear relation between concentration of hydrogen peroxide calculates the mistake in hydrogen peroxide solution to be measured
Hydrogen peroxide concentration.
Embodiment 2:
A kind of detection method of concentration of hydrogen peroxide, process is roughly the same with embodiment 1, and difference is, in step 2,
The horseradish peroxidase solution and 100 μ l neighbour benzene two of 100 μ l are first added into the above-mentioned Hydrogen peroxide standard solution of 100 μ l respectively
Amine aqueous solution at 37 DEG C after hybrid reaction 30min, then is separately added into the carbon quantum dot solution of 100 μ l, and with microsyringe into
Row stirs, and hybrid reaction 1min under room temperature obtains standard sample to be tested system.In step 3, successively by standard sample to be tested system
It is placed in RF-6000 sepectrophotofluorometer, sets excitation wavelength as 380nm, excitation and transmite slit are 5nm, detect institute
The standard sample to be tested of stating ties up to the fluorescence intensity at 442nm and 573nm, obtains the fluorescence spectrum of standard sample to be tested system
Figure, as shown in Figure 1, wherein I442For the fluorescence of carbon dots, I573For the fluorescence of 2,3- diaminophenazine, curve a, b, c, d, e are corresponding
Hydrogen peroxide standard solution concentration be followed successively by 0mol/L, 7.5 × 10-5mol/L、1.5×10-4mol/L、2.25×10-4mol/
L、3×10-4mol/L；Using concentration of hydrogen peroxide as abscissa, I573/I442For ordinate, standard curve is drawn, as shown in Fig. 2,
Establish fluorescence intensity I573/I442Linear relation between concentration of hydrogen peroxide, linear equation I573/I442=0.072
×cHydrogen peroxide+ 0.22, R2=0.997.
The yield of the carbon quantum dot prepared in the present embodiment is 43.5%, using the method for the present embodiment to hydrogen peroxide
Detection is limited to 3 × 10-5mol/L。
Embodiment 3:
A kind of detection method of concentration of hydrogen peroxide, process is roughly the same with embodiment 1, and difference is, in step 1,
In carbon quantum dot preparation, pancreatin is dissolved in ultrapure water, after dispersion, is first put it into magnetizer, in the magnetic field strength of 12T
Lower magnetization treatment 15min, is subsequently placed at 37 DEG C and keeps the temperature 30min, then puts it into super-pressure reactor tank, surpasses under 240MPa
HIGH PRESSURE TREATMENT 5min, finally puts it into reaction kettle again, after reacting 10h at 180 DEG C, takes out, freeze-drying.
The yield of the carbon quantum dot prepared in the present embodiment is 53.1%, using the method for the present embodiment to hydrogen peroxide
Detection is limited to 1.52 × 10-5mol/L。
Although the embodiments of the present invention have been disclosed as above, but its is not only in the description and the implementation listed
With it can be fully applied to various fields suitable for the present invention, for those skilled in the art, can be easily
Realize other modification, therefore without departing from the general concept defined in the claims and the equivalent scope, the present invention is simultaneously unlimited
In specific details and legend shown and described herein.
Claims (8)
1. the detection method of concentration of hydrogen peroxide characterized by comprising
Step 1: preparing the peroxidating of carbon quantum dot solution, horseradish peroxidase solution, o-phenylenediamine solution and various concentration
Hydrogen standard solution；
Step 2: isometric horseradish peroxide is successively added into the Hydrogen peroxide standard solution of same volume various concentration respectively
Compound enzyme solutions, o-phenylenediamine solution and carbon quantum dot solution after hybrid reaction, obtain standard sample to be tested system；
Step 3: detecting the standard sample to be tested ties up to fluorescence intensity at 442nm and 573nm, and establish fluorescence intensity
I573/I442Linear relation between concentration of hydrogen peroxide；
Step 4: it is to be measured molten to prepare the hydrogen peroxide unknown with the volume same concentrations of the Hydrogen peroxide standard solution in step 2
Liquid detects fluorescence intensity of the reactant at 442nm and 573nm, according to glimmering after its method hybrid reaction according to step 2
Luminous intensity I573/I442Linear relation between concentration of hydrogen peroxide calculates the peroxidating in hydrogen peroxide solution to be measured
Hydrogen concentration.
2. the detection method of concentration of hydrogen peroxide as described in claim 1, which is characterized in that in step 1, prepare carbon quantum
Point solution, horseradish peroxidase solution, o-phenylenediamine solution and various concentration Hydrogen peroxide standard solution when, use it is molten
Agent is Tris-HCl buffer solution, pH 7.4, concentration 0.05mol/L.
3. the detection method of concentration of hydrogen peroxide as claimed in claim 2, which is characterized in that in step 1, various concentration
The preparation method of Hydrogen peroxide standard solution are as follows: the hydrogen peroxide stoste for being first 10mol/L with pure hydrogen peroxide compound concentration, it will
The stoste carries out gradient dilution with the Tris-HCl buffer solution to get the Hydrogen peroxide standard of the various concentration is molten
The concentration of liquid is followed successively by 0mol/L, 7.5 × 10-5mol/L、1.5×10-4mol/L、2.25×10-4mol/L、3×10-4mol/L。
4. the detection method of concentration of hydrogen peroxide as claimed in claim 2, which is characterized in that in step 1, carbon quantum dot is molten
The concentration of liquid is 5.3 × 10-6G/mL, the concentration of horseradish peroxidase solution are 1 × 10-5G/mL, o-phenylenediamine solution it is dense
Degree is 3 × 10-2mol/L。
5. the detection method of concentration of hydrogen peroxide as described in claim 1, which is characterized in that in step 1, carbon quantum dot
The preparation method comprises the following steps: the pancreatin of 2 parts by weight is dissolved in the ultrapure water of 10 parts by weight, after dispersion, put it into reaction kettle, in
At 180 DEG C react 10h after, take out, freeze-drying to get.
6. the detection method of concentration of hydrogen peroxide as described in claim 1, which is characterized in that in step 2 specifically: first divide
Isometric horseradish peroxidase solution and adjacent benzene are not added into the Hydrogen peroxide standard solution of same volume various concentration
Diamine solution at 37 DEG C after hybrid reaction 30min, then is separately added into isometric carbon quantum dot solution, and use micro-sampling
Device is stirred, hybrid reaction 1min under room temperature.
7. the detection method of concentration of hydrogen peroxide as described in claim 1, which is characterized in that in step 3, fluorescence detection
Condition are as follows: excitation wavelength 380nm, excitation and transmite slit are 5nm.
8. the detection method of concentration of hydrogen peroxide as claimed in claim 5, which is characterized in that when prepared by carbon quantum dot, by pancreas
Enzyme is dissolved in ultrapure water, after dispersion, is first put it into magnetizer, the magnetization treatment 15min under the magnetic field strength of 12T, then
It is placed at 37 DEG C and keeps the temperature 30min, then put it into super-pressure reactor tank, the ultra high pressure treatment 5min under 240MPa, finally again
It puts it into reaction kettle, after reacting 10h at 180 DEG C, takes out, freeze-drying.
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Cited By (5)
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| CN110423773A (en) * | 2019-08-07 | 2019-11-08 | 上海合森生物科技有限公司 | A kind of hydrogen peroxide whole-cell biological sensor and its preparation and application |
| CN110726707A (en) * | 2019-10-30 | 2020-01-24 | 南京医科大学 | Based on N-Ti3C2Composite nano probe of QDs and o-phenylenediamine oxide and ratiometric fluorescence detection method thereof |
| CN110887836A (en) * | 2019-12-13 | 2020-03-17 | 南京师范大学常州创新发展研究院 | Method for detecting content of hydrogen peroxide |
| CN111337467A (en) * | 2020-04-22 | 2020-06-26 | 江西理工大学 | Tetravalent cerium ion fluorescence detection reagent and fluorescence detection method thereof |
| CN113533283A (en) * | 2021-07-15 | 2021-10-22 | 广州珠矶科技有限公司 | Method for detecting hydrogen peroxide by using carbon quantum dots |
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| CN113533283A (en) * | 2021-07-15 | 2021-10-22 | 广州珠矶科技有限公司 | Method for detecting hydrogen peroxide by using carbon quantum dots |
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