CN103728287B - Nano cupric oxide Mimetic enzyme measures the fluorescence analysis method of glucose - Google Patents
Nano cupric oxide Mimetic enzyme measures the fluorescence analysis method of glucose Download PDFInfo
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Abstract
The present invention discloses the fluorescence analysis method </b> that a kind of <b> nano cupric oxide Mimetic enzyme measures glucose, described nano cupric oxide is the fluorescence analysis method that Mimetic enzyme measures glucose, first its feature is by glucose, glucose oxidase and phosphate buffer three mix temperature bath, then add phosphate buffer wherein, terephthalic acid (TPA) and nano cupric oxide continue temperature bath.Fluorescence exciting wavelength and the emission wavelength of the reaction product of above-mentioned mixed liquor are respectively 315nm and 421nm.Glucose content setting-out line scope is 3 ~ 120 μm of ol/L, detects and is limited to 1.94 μm of ol/L.The method can be used for the mensuration of blood sugar concentration.
Description
Technical field
The present invention relates to nano cupric oxide measures glucose fluorescence analysis method as Mimetic enzyme, belong to analytical chemistry and field of nanometer technology.
Background technology
At present, the method for clinical assays blood sugar has ortho-aminotoluene method and enzyme process.Ortho-aminotoluene method is simple to operate, and cost is low, but specificity is poor, and ortho-aminotoluene reagent is unstable, and toxicity is large.Enzyme process utilizes the high selectivity catalysis of enzyme to carry out the method analyzed.According to the difference of enzyme class, enzyme process can be divided into hexokinase method, glucose dehydrogenase method and glucose oxidase-peroxidase method.Mostly be single step reaction in commercial blood glucose meter to complete, but selectivity is poor, by material serious interference such as maltose, wood sugar, galactoses, often causes the false rising of blood sugar detection value; Glucose oxidase-peroxidase method is easy and simple to handle, and the wide and low price of reagent source is the determination method of blood sugar that the Ministry of Public Health of China is recommended.
Fluorescence, refers to a kind of chemiluminescence phenomenon of photoluminescence.When the incident light (normally ultraviolet or X ray) of certain normal temperature material through certain wavelength irradiates, after absorbing luminous energy, enter excited state, and de excitation is sent out and is sent the emergent light longer than the wavelength of incident light immediately; And once stop incident light, luminescence phenomenon also disappears immediately thereupon.The emergent light with this character is just referred to as fluorescence.In recent years because fluorometry has highly sensitive, the range of linearity is wide, and analysis cost is low, and equipment operating is simple and provide advantages such as containing much information, has attracted the extensive concern of people in fields such as analysing of Fen ﹑ Huan border Ke ﹑ clinical medicine.
The present invention is based on the Mimetic enzyme performance that nano cupric oxide is good, produce the reaction of hydrogen peroxide in conjunction with glucose oxidase glucose, provide a kind of nano cupric oxide catalyzing hydrogen peroxide to be oxidized terephthalic acid (TPA) and produce the new method that fluorescence signal measures glucose.
Summary of the invention
The object of the invention is based on the good Mimetic enzyme performance of nano cupric oxide, produce the process of hydrogen peroxide in conjunction with glucose oxidase catalytic oxidation of glucose, provide a kind of nano cupric oxide catalyzing hydrogen peroxide to be oxidized terephthalic acid (TPA) and produce the new method that fluorescence signal measures glucose.
To achieve these goals, the present invention is by the following technical solutions: described a kind of nano cupric oxide is the fluorescence analysis method that Mimetic enzyme measures glucose, it is characterized in that first glucose, glucose oxidase and phosphate buffer three being mixed temperature bath carries out oenoxydase enzymatic reaction, then phosphate buffer is added wherein, terephthalic acid (TPA) and nano cupric oxide continue temperature bath and carry out fluorescence reaction, by the fluorescence intensity of the above-mentioned mixed liquor reaction product of fluorescent spectrophotometer assay.
Maximum excitation wavelength and the emission wavelength of described fluorescence reaction product are respectively 315nm and 421nm.
Described glucose oxidase enzymatic reaction system pH value is preferably 7.0, and glucose oxidase enzyme concentration is preferably 15U/mL.
Described glucose oxidase enzymatic reaction temperature is preferably 37 DEG C, and the reaction time is preferably 20 minutes.
The pH value of described fluorescence reaction system is preferably 7.0, and nano cupric oxide concentration is preferably 0.4mg/L, and P-phthalic acid at concentration is preferably 3.0mmol/L.
Described fluorescence reaction temperature is preferably 45 DEG C, and the reaction time is preferably 120 minutes.
The nano cupric oxide that utilizes of the present invention is for the fluorescence analysis method of Mimetic enzyme mensuration glucose, it is characterized in that in EP pipe, add 62.5 μ L concentration is respectively the glucose oxidase of 150U/mL, 62.5 μ L concentration are 200mmol/L, the phosphate buffered solution of pH7.0 and the glucose solution of 0.5mL variable concentrations and form the mixed liquor of different concentration of glucose, the mixed liquor of above-mentioned different concentration of glucose is placed in respectively 37 DEG C of temperature baths, after 20 minutes, 3.525mL concentration is the phosphate buffer (pH7.0) of 200mmol/L wherein, 0.8mL concentration is the terephthalic acid (TPA) of 18.75mmol/L and 50 μ L concentration is the nano cupric oxide of 40mg/L, 45 DEG C of temperature baths, its fluorescence intensity at 421nm place is measured respectively after 120 minutes, excitation wavelength is 315nm, with fluorescence intensity, typical curve is obtained for mensuration glucose to concentration of glucose mapping.
Described nano cupric oxide is obtained by following steps: 1) get the Schweinfurt green solution 150ml of 0.02mol/L and 0.5ml glacial acetic acid and join and be equipped with in the three-necked bottle of condenser pipe, be heated with stirring to boiling; 2) add the sodium hydroxide solution 10ml of 0.04g/ml fast, add rear continuation stirring 5 minutes, obtain brown cupric oxide precipitation; 3) the cupric oxide pelleting centrifugation will be obtained by reacting, with absolute ethanol washing three times, drying under reduced pressure, obtains nano-cupric oxide powder.
A kind of nano cupric oxide that utilizes of the present invention measures the fluorescence analysis method of serum glucose level for Mimetic enzyme, it is characterized in that it is made up of following steps: serum, glucose oxidase and the phosphate buffer mixing temperature bath after (1) adds dilution respectively in EP pipe; (2) in above-mentioned EP pipe, add phosphate buffer, terephthalic acid (TPA) and nano cupric oxide continue temperature bath; (3) reaction product is placed in fluorospectrophotometer fluorescence intensity, measures blood sugar concentration according to glucose standard curve.
The nano cupric oxide that utilizes of the present invention is for the fluorescence analysis method of Mimetic enzyme mensuration serum glucose, it is characterized in that in EP pipe, add 62.5 μ L concentration is respectively the glucose oxidase of 150U/mL, 62.5 μ L concentration are 200mmol/L, the phosphate buffered solution of pH7.0 and 0.5mL dilute the serum of 40 times and form mixed liquor, above-mentioned mixed liquor is placed in 37 DEG C of temperature baths, after 20 minutes, 3.525mL concentration is 200mmol/L wherein, pH7.0 phosphate buffer, 0.8mL concentration is the terephthalic acid (TPA) of 18.75mmol/L and 50 μ L concentration is the nano cupric oxide of 40mg/L, 45 DEG C of temperature baths, its fluorescence intensity at 421nm place is measured after 120 minutes, excitation wavelength is 315nm, described nano cupric oxide is obtained by following steps: 1) get the Schweinfurt green solution 150ml of 0.02mol/L and 0.5ml glacial acetic acid and join and be equipped with in the three-necked bottle of condenser pipe, be heated with stirring to boiling, 2) add the sodium hydroxide solution 10ml of 0.04g/ml fast, add rear continuation stirring 5 minutes, obtain brown cupric oxide precipitation, 3) the cupric oxide pelleting centrifugation will be obtained by reacting, with absolute ethanol washing three times, drying under reduced pressure, obtains nano-cupric oxide powder.
Technical scheme concrete steps of the present invention are as follows:
(1) preparation of nano cupric oxide:
Get Schweinfurt green solution and glacial acetic acid to join and be equipped with in the three-necked bottle of condenser pipe, be heated with stirring to boiling, add sodium hydroxide solution fast, after adding, after continuing to stir, obtain cupric oxide.By centrifugal immediately for the cupric oxide be obtained by reacting, with absolute ethanol washing, drying under reduced pressure, obtain nano-cupric oxide powder.Nano-cupric oxide powder is scattered in redistilled water and obtains brown nano cupric oxide colloidal solution.
The concrete preparation process of nano cupric oxide is as follows:
(1) get the Schweinfurt green solution 150ml of 0.02mol/L and 0.5ml glacial acetic acid to join and be equipped with in the three-necked bottle of condenser pipe, be heated with stirring to boiling;
(2) add the sodium hydroxide solution 10ml of 0.04g/ml fast, add rear continuation stirring 5 minutes, obtain brown cupric oxide precipitation;
(3) the cupric oxide pelleting centrifugation will be obtained by reacting, with absolute ethanol washing three times, drying under reduced pressure, obtains the nano-cupric oxide powder that diameter is 6nm.
(2) mensuration of glucose
In EP pipe, add the glucose solution of glucose oxidase, phosphate buffered solution and variable concentrations respectively, mixed liquor is put into 37 DEG C of thermostat water baths and leave standstill.Add phosphate buffer wherein after 20 minutes, terephthalic acid (TPA) and nano cupric oxide continue temperature bath, and bath temperature is 45 DEG C.Its fluorescence intensity is measured after 120 minutes.With fluorescence intensity concentration of glucose mapped and obtain typical curve.
(3) mensuration of blood sugar
Glucose is replaced with the blood serum sample after dilution and repeat step 2, gained fluorescence intensity is substituted into the mensuration that typical curve can carry out blood sugar.
advantage of the present invention:
The present invention utilizes the Mimetic enzyme characteristic of nano cupric oxide, with the reaction bonded of glucose oxidase glucose, successfully constructs a kind of fluorescence analysis method detecting glucose.The range of linearity that this technology measures glucose is 3 ~ 120 μm of ol/L, and its detection is limited to 1.94 μm of ol/L.The present invention has highly sensitive, and sample requirements is few, favorable reproducibility, low cost and other advantages.The successful mensuration of serum glucose sample shows the method in clinical detection, has good potentiality in the practical application such as food inspection and environmental monitoring.
Accompanying drawing explanation
Fig. 1 is the effect diagram of pH value to fluorescence intensity.
Fig. 2 is the effect diagram of temperature of reaction to fluorescence intensity.
Fig. 3 is the effect diagram of nano cupric oxide concentration to fluorescence intensity.
Fig. 4 is the effect diagram of P-phthalic acid at concentration to fluorescence intensity.
Fig. 5 is the effect diagram of glucose oxidase time of enzymatic reacting to fluorescence intensity.
Fig. 6 is the effect diagram of fluorescence reaction time to fluorescence intensity.
Fig. 7 is the canonical plotting of glucose.
Fig. 8 is lactose, maltose, the fluorescence intensity comparison diagram of fructose and glucose.
Embodiment
example 1:
The concrete preparation process of nano cupric oxide is as follows: (1) is got the Schweinfurt green solution 150ml of 0.02mol/L and 0.5ml glacial acetic acid and joined and be equipped with in the three-necked bottle of condenser pipe, is heated with stirring to boiling; (2) add the sodium hydroxide solution 10ml of 0.04g/ml fast, add rear continuation stirring 5 minutes, obtain brown cupric oxide precipitation; (3) the cupric oxide pelleting centrifugation will be obtained by reacting, with absolute ethanol washing three times, drying under reduced pressure, obtains the nano-cupric oxide powder that diameter is 6nm.
example 2:
Be the terephthalic acid (TPA) of 18.75mmol/L by 0.8mL concentration, nano cupric oxide prepared by the example 1 of 0.5mL concentration to be hydrogen peroxide and the 50 μ L concentration of 1mol/L be 40mg/L joins in the phosphate buffer (pH3 ~ 10) that 3.65mL concentration is the different pH of 200mmol/L, mixing shakes up and is placed on 45 DEG C of temperature baths, measures its fluorescence intensity at 421nm place (excitation wavelength is 315nm) after 20 minutes.As shown in Figure 1, fluorescence intensity reaches maximal value when pH is 7.0.
example 3:
Be the terephthalic acid (TPA) of 18.75mmol/L by 0.8mL concentration, nano cupric oxide prepared by the example 1 of 0.5mL concentration to be the hydrogen peroxide of 1mol/L and 50 μ L concentration be 40mg/L joins in the phosphate buffer (pH7.0) that 3.65mL concentration is 200mmol/L, mixing shakes up and is placed on the bath of different temperatures (20 ~ 55 DEG C) temperature, measures its fluorescence intensity at 421nm place (excitation wavelength is 315nm) after 20 minutes.As shown in Figure 2, fluorescence intensity reaches maximal value 45 DEG C time.
example 4:
Be the terephthalic acid (TPA) of 18.75mmol/L by 0.8mL concentration, 0.5mL concentration is that nano cupric oxide (0 ~ 80mg/L) prepared by the hydrogen peroxide of 1mol/L and the example 1 of 50 μ L variable concentrations joins in the phosphate buffer (pH7.0) that 3.65mL concentration is 200mmol/L, mixing shakes up and is placed on 45 DEG C of temperature baths, measures its fluorescence intensity at 421nm place (excitation wavelength is 315nm) after 20 minutes.As shown in Figure 3, fluorescence intensity increases with nano cupric oxide concentration in mixed liquor and increases and reach maximum when concentration is 0.4 ~ 0.8mg/L.
example 5:
By the terephthalic acid (TPA) (0 ~ 22.5mmol/L) of 0.8mL variable concentrations, nano cupric oxide prepared by the example 1 of 0.5mL concentration to be the hydrogen peroxide of 1mol/L and 50 μ L concentration be 40mg/L joins in the phosphate buffer (pH7.0) that 3.65mL concentration is 200mmol/L, mixing shakes up and is placed on 45 DEG C of temperature baths, measures its fluorescence intensity at 421nm place (excitation wavelength is 315nm) after 20 minutes.As shown in Figure 4, fluorescence intensity increases with P-phthalic acid at concentration in mixed liquor and increases, and when final concentration is 3.0mmol/L, fluorescence intensity reaches stationary value.
example 6:
In EP pipe, add 62.5 μ L concentration is respectively the glucose oxidase of 150U/mL, the glucose solution of 62.5 μ L concentration to be 200mmol/L phosphate buffered solution (pH7.0) and 0.5mL concentration be 4mmol/L forms mixed liquor, above-mentioned mixed liquor is placed in 37 DEG C of temperature baths, the phosphate buffer (pH7.0) that 3.525mL concentration is 200mmol/L is added wherein after different time (0-100 minute), nano cupric oxide prepared by the example 1 of 0.8mL concentration to be the terephthalic acid (TPA) of 18.75mmol/L and 50 μ L concentration be 40mg/L, 45 DEG C of temperature baths, its fluorescence intensity at 421nm place (excitation wavelength is 315nm) is measured after 20 minutes.As shown in Figure 5, fluorescence intensity, at first 20 minutes rapid developments, increasess slowly when 20 to 70 minutes, in reaction signal reduction after 70 minutes.
example 7:
In EP pipe, add 62.5 μ L concentration is respectively the glucose oxidase of 150U/mL, the glucose solution of 62.5 μ L concentration to be 200mmol/L phosphate buffered solution (pH7.0) and 0.5mL concentration be 4mmol/L forms mixed liquor, above-mentioned mixed liquor is placed in 37 DEG C of temperature baths, the phosphate buffer (pH7.0) that 3.525mL concentration is 200mmol/L is added wherein after 20 minutes, nano cupric oxide prepared by the example 1 of 0.8mL concentration to be the terephthalic acid (TPA) of 18.75mmol/L and 50 μ L concentration be 40mg/L, 45 DEG C of temperature baths, different time (20-140 minute) measures its fluorescence intensity at 421nm place (excitation wavelength is 315nm) afterwards.As shown in Figure 6, fluorescence intensity increases with the prolongation of fluorescence reaction time, and reaches stationary value after 120 minutes.
example 8:
In EP pipe, add 62.5 μ L concentration is respectively the glucose oxidase of 150U/mL, 62.5 μ L concentration are the mixed liquor that the glucose solution of 200mmol/L phosphate buffered solution (pH7.0) and 0.5mL variable concentrations forms different glucose concentration, the mixed liquor of above-mentioned different glucose concentration is placed in respectively 37 DEG C of temperature baths, the phosphate buffer (pH7.0) that 3.525mL concentration is 200mmol/L is added wherein after 20 minutes, nano cupric oxide prepared by the example 1 of 0.8mL concentration to be the terephthalic acid (TPA) of 18.75mmol/L and 50 μ L concentration be 40mg/L, 45 DEG C of temperature baths, its fluorescence intensity at 421nm place (excitation wavelength is 315nm) is measured respectively after 120 minutes.With fluorescence intensity, typical curve is obtained to concentration of glucose mapping.As shown in Figure 7, fluorescence intensity and concentration of glucose linear within the scope of 3 ~ 120 μm of ol/L, detect be limited to 1.94 μm of ol/L.
example 9:
In EP pipe, add 62.5 μ L concentration is respectively the glucose oxidase of 150U/mL, the glucose solution of 62.5 μ L concentration to be 200mmol/L phosphate buffered solution (pH7.0) and 0.5mL concentration be 1mol/L forms mixed liquor, above-mentioned mixed liquor is placed in 37 DEG C of temperature baths, the phosphate buffer (pH7.0) that 3.525mL concentration is 200mmol/L is added wherein after 20 minutes, nano cupric oxide prepared by the example 1 of 0.8mL concentration to be the terephthalic acid (TPA) of 18.75mmol/L and 50 μ L concentration be 40mg/L, 45 DEG C of temperature baths, its fluorescence intensity at 421nm place (excitation wavelength is 315nm) is measured after 120 minutes.Repeat 6 times, the relative standard deviation of testing result is 1.54%.
example 10:
In EP pipe, add 62.5 μ L concentration is respectively the glucose oxidase of 150U/mL, 62.5 μ L concentration are the different material mixed liquor that 200mmol/L phosphate buffered solution (pH7.0) and 0.5mL different material (lactose of 40mmol/L or 10mmol/L maltose or 10mmol/L fructose) replace glucose solution to be formed, above-mentioned formed different material mixed liquor is placed in respectively 37 DEG C of temperature baths, after 20 minutes, 3.525mL concentration is the phosphate buffer (pH7.0) of 200mmol/L wherein, nano cupric oxide prepared by the example 1 of 0.8mL concentration to be the terephthalic acid (TPA) of 18.75mmol/L and 50 μ L concentration be 40mg/L, 45 DEG C of temperature baths, its fluorescence intensity at 421nm place (excitation wavelength is 315nm) is measured respectively after 120 minutes.As shown in Figure 8, compared with the signal produced with the glucose of 1mmol/L, the signal of the lactose of 40mmol/L or 10mmol/L maltose or the generation of 10mmol/L fructose all can be ignored.
example 11:
In EP pipe, add 62.5 μ L concentration is respectively the glucose oxidase of 150U/mL, 62.5 μ L concentration are that the serum that 200mmol/L phosphate buffered solution (pH7.0) and 0.5mL dilute 40 times forms mixed liquor, above-mentioned mixed liquor is placed in 37 DEG C of temperature baths, the phosphate buffer (pH7.0) that 3.525mL concentration is 200mmol/L is added wherein after 20 minutes, nano cupric oxide prepared by the example 1 of 0.8mL concentration to be the terephthalic acid (TPA) of 18.75mmol/L and 50 μ L concentration be 40mg/L, 45 DEG C of temperature baths, its fluorescence intensity at 421nm place (excitation wavelength is 315nm) is measured after 120 minutes.Glucose content in blood serum sample is calculated, the data consistent that this numerical value and glucose oxidase-horseradish peroxidase-TMB colourimetry obtain through embodiment 8 gained glucose standard curve.The sample tests recovery is 90.7% ~ 105.8%, and relative standard deviation is 0.5 ~ 3.6%, and this method is reliably suitable for as can be seen here.
The foregoing is only preferred embodiment of the present invention, not in order to limit the present invention, all any amendments done within the spirit and principles in the present invention, equivalent replacement and improvement etc., all should be included within protection scope of the present invention.
Claims (5)
1. a nano cupric oxide is the fluorescence analysis method that Mimetic enzyme measures glucose, it is characterized in that first glucose, glucose oxidase and phosphate buffer three being mixed temperature bath carries out oenoxydase enzymatic reaction, then phosphate buffer is added wherein, terephthalic acid (TPA) and nano cupric oxide continue temperature bath and carry out fluorescence reaction, by the fluorescence intensity of the above-mentioned mixed liquor reaction product of fluorescent spectrophotometer assay; The maximum excitation wavelength of fluorescence reaction product and emission wavelength are respectively 315nm and 421nm; Glucose oxidase enzymatic reaction system pH value is 7.0, and glucose oxidase enzyme concentration is 15U/mL; The pH value of fluorescence reaction system is 7.0, and nano cupric oxide concentration is 0.4mg/L, and P-phthalic acid at concentration is 3.0mmol/L; Described nano cupric oxide is obtained by following steps: 1) get the Schweinfurt green solution 150ml of 0.02mol/L and 0.5ml glacial acetic acid and join and be equipped with in the three-necked bottle of condenser pipe, be heated with stirring to boiling; 2) add the sodium hydroxide solution 10ml of 0.04g/ml fast, add rear continuation stirring 5 minutes, obtain cupric oxide precipitation; 3) the cupric oxide pelleting centrifugation will be obtained by reacting, with absolute ethanol washing three times, drying under reduced pressure, obtains nano-cupric oxide powder.
2. nano cupric oxide according to claim 1 is the fluorescence analysis method that Mimetic enzyme measures glucose, and it is characterized in that glucose oxidase enzymatic reaction temperature is 37 DEG C, the reaction time is 20 minutes.
3. nano cupric oxide according to claim 1 and 2 is the fluorescence analysis method that Mimetic enzyme measures glucose, and it is characterized in that fluorescence reaction temperature is 45 DEG C, the reaction time is 120 minutes.
4. one kind utilizes nano cupric oxide for the fluorescence analysis method of Mimetic enzyme mensuration glucose, it is characterized in that in EP pipe, add 62.5 μ L concentration is respectively the glucose oxidase of 150U/mL, 62.5 μ L concentration are 200mmol/L, the phosphate buffered solution of pH7.0 and the glucose solution of 0.5mL variable concentrations and form the mixed liquor of different concentration of glucose, the mixed liquor of above-mentioned different concentration of glucose is placed in respectively 37 DEG C of temperature baths, the phosphate buffer (pH7.0) that 3.525mL concentration is 200mmol/L is added wherein after 20 minutes, 0.8mL concentration is the terephthalic acid (TPA) of 18.75mmol/L and 50 μ L concentration is the nano cupric oxide of 40mg/L, 45 DEG C of temperature baths, its fluorescence intensity at 421nm place is measured respectively after 120 minutes, excitation wavelength is 315nm, with fluorescence intensity, typical curve is obtained for mensuration glucose to concentration of glucose mapping, described nano cupric oxide is obtained by following steps: 1) get the Schweinfurt green solution 150ml of 0.02mol/L and 0.5ml glacial acetic acid and join and be equipped with in the three-necked bottle of condenser pipe, be heated with stirring to boiling, 2) add the sodium hydroxide solution 10ml of 0.04g/ml fast, add rear continuation stirring 5 minutes, obtain cupric oxide precipitation, 3) the cupric oxide pelleting centrifugation will be obtained by reacting, with absolute ethanol washing three times, drying under reduced pressure, obtains nano-cupric oxide powder.
5. utilize nano cupric oxide to measure a fluorescence analysis method for serum glucose level for Mimetic enzyme, it is characterized in that it is made up of following steps: serum, glucose oxidase and the phosphate buffer mixing temperature bath after (1) adds dilution respectively in EP pipe, (2) in above-mentioned EP pipe, add phosphate buffer, terephthalic acid (TPA) and nano cupric oxide continue temperature bath, (3) reaction product is placed in fluorospectrophotometer fluorescence intensity, measures blood sugar concentration according to glucose standard curve, in EP pipe, add 62.5 μ L concentration is respectively the glucose oxidase of 150U/mL, 62.5 μ L concentration are 200mmol/L, the phosphate buffered solution of pH7.0 and 0.5mL dilute the serum of 40 times and form mixed liquor, above-mentioned mixed liquor is placed in 37 DEG C of temperature baths, adding 3.525mL concentration after 20 minutes is wherein 200mmol/L, pH7.0 phosphate buffer, 0.8mL concentration is the terephthalic acid (TPA) of 18.75mmol/L and 50 μ L concentration is the nano cupric oxide of 40mg/L, 45 DEG C of temperature baths, its fluorescence intensity at 421nm place is measured after 120 minutes, excitation wavelength is 315nm, described nano cupric oxide is obtained by following steps: 1) get the Schweinfurt green solution 150ml of 0.02mol/L and 0.5ml glacial acetic acid and join and be equipped with in the three-necked bottle of condenser pipe, be heated with stirring to boiling, 2) add the sodium hydroxide solution 10ml of 0.04g/ml fast, add rear continuation stirring 5 minutes, obtain cupric oxide precipitation, 3) the cupric oxide pelleting centrifugation will be obtained by reacting, with absolute ethanol washing three times, drying under reduced pressure, obtains nano-cupric oxide powder.
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