CN110208232A - The method for quickly measuring ascorbic acid with nano cupric oxide self-activation cascade catalytic fluorometry system - Google Patents
The method for quickly measuring ascorbic acid with nano cupric oxide self-activation cascade catalytic fluorometry system Download PDFInfo
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Abstract
The present invention discloses a kind of method for quickly measuring ascorbic acid with nano cupric oxide self-activation cascade catalytic fluorometry system, it is characterized in that ascorbic acid, after nano cupric oxide and terephthalic acid (TPA) three mix warm bath, its fluorescence exciting wavelength of reaction product and launch wavelength are respectively 315 nm and 421 nm.Transmitting fluorescence intensity is detected through sepectrophotofluorometer.Nano cupric oxide is added in terephthalic acid (TPA)-ascorbic acid system can significantly increase fluorescence signal.Using terephthalic acid (TPA)-ascorbic acid-nano cupric oxide fluorescence system can Ascorbic Acid content be measured, the range of linearity be 7.5 × 10‑7‑7.5×10‑6M and 1.25 × 10‑5‑1.25×10‑4M, detection are limited to 2.92 × 10‑8 M。
Description
Technical field
Hydrogen peroxide, which is generated, as the oxidase catalyzed Ascorbic Acid Oxidation of simulation the present invention relates to nano cupric oxide is coupled it
The method that Catalyzed Synthesis By Peroxidase aoxidizes terephthalic acid (TPA) enhancing fluoremetry ascorbic acid, belongs to analytical chemistry and nanotechnology
Field.
Background technique
Ascorbic acid (ascorbic acid, AA) is prevalent in fresh fruit and vegetables, is had anti-oxidant and is improved
The effect of immunity of organisms can participate in internal a series of metabolism and redox reaction, increase to the resistance for infecting disease.
Clinical research shows that the number of AA content in the generation and human body of some diseases has substantial connection.Research for ascorbic acid
Early oneself causes the common concern of people, and detection method has electrochemical process, fluorescence method, chromatography, chemiluminometry, efficient liquid
Phase chromatography etc..
In recent years, the unique simulation enzymatic property of nano material has been widely used in the neck such as environment measuring and biomedicine
Domain.But the catalytic performance of these biomimetic materials is combined and is built into one similar to the function that common are tissue in organism
The enzyme-linked system (artificial enzyme's cascade system) of energy is still great difficulty and challenge.This artificial enzyme's cascade system has huge
Big potentiality and advantage will effectively eliminate string if successfully the analogue enztme group of different function can be installed in same system
Between connection reaction because reaction condition it is different caused by incompatibility.
Fluorescence analysis has high sensitivity, and the range of linearity is wide, and analysis cost is low, and equipment operation is simple and provides information
It measures the advantages that big, has attracted the extensive concern of people in fields such as the border Fen analysisization ﹑ Huan Ke ﹑ clinical medicine.Nanometer material
Expect specific surface area great ﹑ adsorptivity Qiang ﹑ Shui Rong ﹑ high activity and it is highly selective many advantages, such as, nano material is extensive in recent years
It is applied in fluorescence analysis method.And the combination of nanotechnology will undoubtedly open up in terms of various actual analysis for fluorescence analysis
More extensive application prospect.
Oxidase catalyzed Ascorbic Acid Oxidation generation hydrogen peroxide, which is simulated, the present invention is based on nano cupric oxide is coupled its peroxide
Compound enzymatic oxidation terephthalic acid (TPA) enhances the process of fluorescence, provides a kind of fluorescence analysis side of quickly measurement ascorbic acid
Method.
Summary of the invention
The purpose of the present invention is being based on the good catalytic performance of nano cupric oxide, a kind of catalysis Ascorbic Acid Oxidation pair is provided
The new method of phthalic acid enhancing fluorescence signal.
To achieve the goals above, the invention adopts the following technical scheme:
A method of ascorbic acid quickly being measured with nano cupric oxide self-activation cascade catalytic fluorometry system, it is characterized in that anti-bad
Hematic acid, nano cupric oxide and terephthalic acid (TPA) three mix warm bath, its fluorescence exciting wavelength of reaction product and launch wavelength difference
For 315 nm and 421 nm；Transmitting fluorescence intensity is detected through sepectrophotofluorometer, and nano cupric oxide can significantly increase fluorescence letter
Number intensity, intensification factor is about 41 times.
The method for quickly measuring ascorbic acid with nano cupric oxide self-activation cascade catalytic fluorometry system, feature
The pH value for being reaction system is 8.0.
The method for quickly measuring ascorbic acid with nano cupric oxide self-activation cascade catalytic fluorometry system, feature
It is that bath temperature is 45 DEG C in reaction system, the reaction time is 10 minutes.
The method for quickly measuring ascorbic acid with nano cupric oxide self-activation cascade catalytic fluorometry system, feature
The concentration for being nano cupric oxide in reaction system is 1.25 mg/L.
The method for quickly measuring ascorbic acid with nano cupric oxide self-activation cascade catalytic fluorometry system, feature
It is P-phthalic acid at concentration in reaction system is 2.5 mmol/L.
It is of the present invention it is a kind of with nano cupric oxide self-activation cascade catalytic fluorometry system quickly measure ascorbic acid
Method, it is characterized in that be separately added into EP pipe the ascorbic acid solution of various concentration, phosphate buffer, terephthalic acid (TPA),
Nano cupric oxide, mixed liquor warm bath, reaction product is put into sepectrophotofluorometer and detects emitted luminescence intensity.
The method for quickly measuring ascorbic acid with nano cupric oxide self-activation cascade catalytic fluorometry system, feature
The ascorbic acid solution volume for being various concentration be added is 0.5 ml, and the phosphate buffer volume of addition is 2.95 ml,
Concentration is 200 mmol/L, and 8.0,20 mmol/L terephthalic acid (TPA) volume of pH is the nano oxidized of 0.5 ml and 100 mg/L
Copper volume is 50 μ l, mixed liquor warm bath at 45 DEG C, and product fluorescence intensity is through fluorescent spectrophotometer assay after ten minutes, with
Fluorescence intensity Ascorbic Acid concentration maps to obtain standard curve, and fluorescence intensity and ascorbic acid concentrations are 7.5 × 10-7-7.5×
10-6M and 1.25 × 10-5-1.25×10-4In a linear relationship within the scope of M, detection is limited to 2.92 × 10-8 M。
It is of the present invention it is a kind of with nano cupric oxide self-activation cascade catalytic fluorometry system quickly measure ascorbic acid
Method, it is characterized in that steps are as follows: (1) drug being added to ultrasonic dissolution in distilled water, drug sample solution is obtained after dilution；
Orange juice directly dilutes to obtain juice samples solution；(2) by drug sample solution or juice samples solution, nano cupric oxide and to benzene two
Formic acid three mixes warm bath；(3) reaction product is placed in fluorescence intensity in sepectrophotofluorometer, according to ascorbic acid
Linearity curve calculates drug or juice samples content.
The method for quickly measuring ascorbic acid with nano cupric oxide self-activation cascade catalytic fluorometry system, feature
Be: (1) drug is added to ultrasonic dissolution in 50 mL distilled water, and 40 times of dilution obtains drug sample solution after standing；Orange juice is direct
40 times of dilution obtain juice samples solution；(2) by 0.5 mL drug sample solution or juice samples solution, volume is 0.5 ml concentration
To be added to volume be 2.95 ml to the nano cupric oxide for being 50 μ l for 20 mmol/L terephthalic acid (TPA)s and 100 mg/L volumes, dense
Degree is 200 mmol/L, mixes and is placed in 45 DEG C of water-baths 10 minutes in the phosphate buffer that pH is 8.0；(3) reaction is produced
Object is placed in fluorescence intensity in sepectrophotofluorometer, is measured according to the linearity curve of ascorbic acid anti-bad in drug or orange juice
Hematic acid content.
The method for quickly measuring ascorbic acid with nano cupric oxide self-activation cascade catalytic fluorometry system, feature
It is that nano cupric oxide is specifically prepared by following step: (1) takes 150 ml of acetic acid copper solution and 0.5 ml ice of 0.02 mol/L
Acetic acid is added in the three-necked bottle equipped with condenser pipe, is heated with stirring to boiling；(2) sodium hydroxide of 0.04 g/ml is rapidly joined
10 ml of solution continues stirring 5 minutes after adding, obtain brown copper oxide precipitating；(3) cupric oxide that reaction obtains is sunk
It forms sediment and is centrifuged, washed with dehydrated alcohol and be dried under reduced pressure the nano-cupric oxide powder for being 6 nm to get diameter three times, it will be nano oxidized
Copper powder body, which is scattered in secondary distilled water, obtains brown nano cupric oxide colloidal solution.
Specifically, the present invention adopts the following technical scheme:
The method for quickly measuring ascorbic acid with nano cupric oxide self-activation cascade catalytic fluorometry system, it is characterized in that anti-
After bad hematic acid, nano cupric oxide and terephthalic acid (TPA) three mix warm bath, its fluorescence exciting wavelength of reaction product and launch wavelength
Respectively 315 nm and 421 nm.Transmitting fluorescence intensity is detected through sepectrophotofluorometer.
The method for quickly measuring ascorbic acid with nano cupric oxide self-activation cascade catalytic fluorometry system, feature
The pH value for being reaction system is 8.0.
The method for quickly measuring ascorbic acid with nano cupric oxide self-activation cascade catalytic fluorometry system, feature
Be reaction temperature be 45 DEG C.
The method for quickly measuring ascorbic acid with nano cupric oxide self-activation cascade catalytic fluorometry system, feature
Being is 10 minutes in the reaction time.
The method for quickly measuring ascorbic acid with nano cupric oxide self-activation cascade catalytic fluorometry system, feature
The nano oxidized copper concentration for being reaction system is 1.25 mg/L.
The method for quickly measuring ascorbic acid with nano cupric oxide self-activation cascade catalytic fluorometry system, feature
The P-phthalic acid at concentration for being reaction system is 2.5 mmol/L.
The method for quickly measuring ascorbic acid with nano cupric oxide self-activation cascade catalytic fluorometry system, feature
Be to be made of following steps: being separately added into the ascorbic acid of various concentration in EP pipe, phosphate buffer, terephthalic acid (TPA),
Reaction product is put into sepectrophotofluorometer after mixed liquor warm bath and detects emitted luminescence intensity by nano cupric oxide.
The method for quickly measuring ascorbic acid with nano cupric oxide self-activation cascade catalytic fluorometry system, feature
The volume for being various concentration ascorbic acid be added is 0.5 ml；The phosphate buffer volume of addition is 2.95 mL, concentration
For 200 mmol/L, pH 8.0；The terephthalic acid (TPA) volume of addition is 0.5 mL, and concentration is 20 mmol/L；The nanometer of addition
Copper oxide volume is 50 μ L, and concentration is 100 mg/L.Mixed liquor warm bath at 45 DEG C after ten minutes, measures it at 421 nm
Fluorescence intensity (excitation wavelength be 315 nm).
Specific step is as follows for technical solution of the present invention:
(1) preparation of nano cupric oxide:
It takes acetic acid copper solution and glacial acetic acid to be added in the three-necked bottle equipped with condenser pipe, is heated with stirring to boiling, rapidly joins hydrogen
Sodium hydroxide solution after adding, after continuing stirring, obtains cupric oxide.The cupric oxide that reaction obtains is centrifuged immediately, is used
Dehydrated alcohol washing, is dried under reduced pressure to get nano-cupric oxide powder.It disperses nano-cupric oxide powder in secondary distilled water and obtains
To brown nano cupric oxide colloidal solution.
Specific preparation process is as follows for nano cupric oxide:
(1) 150 ml of acetic acid copper solution and 0.5 ml glacial acetic acid for taking 0.02 mol/L are added to the three-necked bottle equipped with condenser pipe
In, it is heated with stirring to boiling；
(2) 10 ml of sodium hydroxide solution for rapidly joining 0.04 g/ml continues stirring 5 minutes after adding, obtain brown oxidation
Copper precipitating；
(3) the cupric oxide precipitating centrifugation obtained reaction, is washed three times with dehydrated alcohol, is dried under reduced pressure to get nano oxygen
Change copper powder body.
(2) nano cupric oxide catalysis Ascorbic Acid Oxidation terephthalic acid (TPA) enhances fluorescence
Phosphate buffer, terephthalic acid (TPA), nano cupric oxide and ascorbic acid are mixed and shaken up.Mixed liquor is placed in 45 DEG C of temperature
Bath after ten minutes, measures its fluorescence intensity at 421 nm (excitation wavelength is 315 nm).
(3) measurement of ascorbic acid
The ascorbic acid of phosphate buffer, terephthalic acid (TPA), nano cupric oxide and various concentration is mixed and is shaken up.Mixed liquor is set
After ten minutes in 45 DEG C of warm bath, measure its fluorescence intensity at 421 nm (excitation wavelength is 315 nm).With fluorescence intensity pair
Ascorbic acid concentrations map to obtain standard curve.
(the measurement of ascorbic acid in four) Yao Pin ﹑ orange juices
Ascorbic acid is replaced with into the Yao Pin ﹑ juice samples after dilution and repeats step 3, it is bent that gained fluorescence intensity is substituted into standard
Line can carry out the measurement of ascorbic acid content in drug.
Advantages of the present invention:
(1) present invention successfully constructs a kind of ascorbic acid detection architecture, and nano cupric oxide is catalyzed Ascorbic Acid Oxidation to benzene two
Formic acid enhances fluorescence signal, shows the good simulation oxidizing ferment-peroxidase catalytic performance of nano cupric oxide.
(2) nano cupric oxide preparation process used in the present invention is simple and quick.
(3) present invention is low to the processing requirement of sample, and drug sample only needs to use distilled water ultrasonic dissolution, can examine after dilution
It surveys.
(4) detection speed of the invention is fast, and the pretreatment and detection of practical drug sample can be completed in 20 minutes.
(5) range of linearity of technology measurement ascorbic acid is 7.5 × 10-7-7.5×10-6M and 1.25 × 10-5-1.25
×10-4M, detection are limited to 2.92 × 10-8M.The present invention has high sensitivity, and high specificity, favorable reproducibility is at low cost etc.
Advantage.This method will show unlimited potentiality in clinical detection, the practical applications such as food and medicine analysis and environmental monitoring.
Detailed description of the invention
Fig. 1 is nano cupric oxide-terephthalic acid (TPA)-ascorbic acid system Fluorescence Enhancement figure, a, TA in Fig. 1; b,
TA+CuO NPs; c,TA+AA; d, TA+ AA+CuO NPs。
Fig. 2 is pH value to terephthalic acid (TPA)-ascorbic acid-nano cupric oxide fluorescence system influence diagram.
Fig. 3 is nano oxidized copper concentration to terephthalic acid (TPA)-ascorbic acid-nano cupric oxide fluorescence system influence diagram.
Fig. 4 is P-phthalic acid at concentration to terephthalic acid (TPA)-ascorbic acid-nano cupric oxide fluorescence system influence diagram.
Fig. 5 is ascorbic acid concentrations to terephthalic acid (TPA)-ascorbic acid-nano cupric oxide fluorescence system influence diagram.
Fig. 6 is temperature to terephthalic acid (TPA)-ascorbic acid-nano cupric oxide fluorescence system influence diagram.
Fig. 7 is the reaction time to terephthalic acid (TPA)-ascorbic acid-nano cupric oxide fluorescence system influence diagram.
Fig. 8 is terephthalic acid (TPA)-ascorbic acid-nano cupric oxide fluorescence system of determination ascorbic acid canonical plotting.
Fig. 9 is the fluorescence intensity comparison diagram of various carbohydrates, amino acids, acids and ascorbic acid, 1 in figure, gluathione
Peptide;2, glycine;3, cysteine;4, uric acid;5, citric acid;6, glucose;7, fructose;8, lactose;
9, maltose;10, sodium chloride;11, oxalic acid;12, alanine;13, ethyl alcohol;14, ascorbic acid.
Specific embodiment
Example 1:
The specific preparation step of nano cupric oxide: (1) 150 ml of acetic acid copper solution of 0.02 mol/L and 0.5 ml glacial acetic acid is taken to add
Enter into the three-necked bottle equipped with condenser pipe, is heated with stirring to boiling；(2) sodium hydroxide solution 10 of 0.04 g/ml is rapidly joined
Ml continues stirring 5 minutes after adding, obtain brown copper oxide precipitating；(3) the cupric oxide precipitating centrifugation obtained reaction,
It is washed three times, is dried under reduced pressure with dehydrated alcohol, nano-cupric oxide powder.
Example 2:
The terephthalic acid (TPA) for being 20 mmol/L by 0.5 mL concentration, 0.5 mL concentration are the ascorbic acid and 50 of 3.0 mmol/L
μ L concentration is that be added to the pH that 2.95 mL concentration are 200 mmol/L be 8.0 to nano cupric oxide made from 100 mg/L examples 1
Phosphate buffer in, mixing shakes up and is placed on 45 DEG C of warm bath, measures its fluorescence intensity at 421 nm after ten minutes
(excitation wavelength is 315 nm).Experiment contrast pipe replaces nano cupric oxide with distilled water.It has detected simultaneously and only exists terephthaldehyde
Acid and with distilled water replace ascorbic acid two groups of experiment contrast pipes fluorescence intensity as shown in Figure 1, nano cupric oxide can be significant
Enhance fluorescence signal intensity, intensification factor is about 41 times.
Example 3:
The terephthalic acid (TPA) for being 20 mmol/L by 0.5 mL concentration, 0.5 mL concentration are the ascorbic acid and 50 μ L of 3 mmol/L
Concentration is that nano cupric oxide made from 100 mg/L examples 1 is added to the different pH value that 2.95 mL concentration are 200 mmol/L
In phosphate buffer (pH 4-9), mixing, which shakes up, is placed on 45 DEG C of warm bath, measures its fluorescence at 421 nm after ten minutes
Intensity (excitation wavelength is 315 nm).As shown in Fig. 2, fluorescence intensity increases with pH value increase and reaches peak when pH is 8.0
Value continues increase pH fluorescence intensity and then reduces.
Example 4:
The terephthalic acid (TPA) for being 20 mmol/L by 0.5 mL concentration, 0.5 mL concentration are the ascorbic acid and 50 of 3.0 mmol/L
Nano cupric oxide (0-200 mg/L) made from μ L various concentration example 1 is added to the pH that 2.95 mL concentration are 200 mmol/L
45 DEG C of warm bath are placed in 8.0 phosphate buffer, mixing is shaken up, measure its fluorescence at 421 nm after ten minutes
Intensity (excitation wavelength is 315 nm).As shown in figure 3, fluorescence intensity increases with copper concentration nano oxidized in mixed liquor and increases simultaneously
Enter plateau when concentration is 1.25-2.5 mg/L.
Example 5:
By the terephthalic acid (TPA) (0-30 mmol/L) of 0.5 mL various concentration, 0.5 mL concentration is the Vitamin C of 3.0 mmol/L
Acid and 50 μ L concentration are that be added to 2.95 mL concentration be 200 mmol/L to nano cupric oxide made from 100 mg/L examples 1
In the phosphate buffer that pH is 8.0, mixing, which shakes up, is placed on 45 DEG C of warm bath, and it is glimmering at 421 nm to measure it after ten minutes
Luminous intensity (excitation wavelength is 315 nm).As shown in figure 4, fluorescence intensity increases with P-phthalic acid at concentration in mixed liquor and is increased
Greatly, in final concentration of 2.5 mmol/L, fluorescence intensity reaches stationary value.
Example 6:
The terephthalic acid (TPA) for being 20 mmol/L by 0.5 mL concentration, the ascorbic acid (0-5 mmol/L) of 0.5 mL various concentration
It is the pH that nano cupric oxide made from 100 mg/L examples 1 is added to that 2.95 mL concentration are 200 mmol/L with 50 μ L concentration
45 DEG C of warm bath are placed in 8.0 phosphate buffer, mixing is shaken up, measure its fluorescence at 421 nm after ten minutes
Intensity (excitation wavelength is 315 nm).As shown in figure 5, fluorescence intensity increases with ascorbic acid concentrations in mixed liquor and is increased,
When final concentration of 0.375 mmol/L, fluorescence intensity reaches maximum.
Example 7:
The terephthalic acid (TPA) for being 20 mmol/L by 0.5 mL concentration, 0.5 mL concentration are the ascorbic acid and 50 of 3.0 mmol/L
μ L concentration is that be added to the pH that 2.95 mL concentration are 200 mmol/L be 8.0 to nano cupric oxide made from 100 mg/L examples 1
Phosphate buffer in, mixing shakes up and is placed on different temperatures (25 ~ 60 DEG C) warm bath, measures it after ten minutes in 421 nm
The fluorescence intensity (excitation wavelength is 315 nm) at place.As shown in fig. 6, fluorescence intensity reaches stationary value at 45-60 DEG C.
Example 8:
The terephthalic acid (TPA) for being 20 mmol/L by 0.5 mL concentration, 0.5 mL concentration are the ascorbic acid and 50 of 3.0 mmol/L
μ L concentration is that be added to the pH that 2.95 mL concentration are 200 mmol/L be 8.0 to nano cupric oxide made from 100 mg/L examples 1
Phosphate buffer in, mixing shakes up and is placed on 45 DEG C of warm bath, and different time (1 ~ 20 minute) measures it at 421 nm afterwards
Fluorescence intensity (excitation wavelength be 315 nm).As shown in fig. 7, fluorescence intensity is to reach stable after ten minutes in the reaction time
Value.
Example 9:
The terephthalic acid (TPA) for being 20 mmol/L by 0.5 mL concentration, the ascorbic acid of 0.5 mL various concentration and 50 μ L concentration are
Nano cupric oxide made from 100 mg/L examples 1 is added to the phosphate that the pH that 2.95 mL concentration are 200 mmol/L is 8.0
In buffer, mixing, which shakes up, is placed on 45 DEG C of warm bath, measures its fluorescence intensity (excitation wavelength at 421 nm after ten minutes
For 315 nm).It maps to obtain standard curve with fluorescence intensity Ascorbic Acid concentration.As shown in figure 8, fluorescence intensity and Vitamin C
Acid concentration is 7.5 × 10-7-7.5×10-6M and 1.25 × 10-5-1.25×10-4In a linear relationship within the scope of M, detection is limited to
2.92×10-8 M。
Example 10:
The terephthalic acid (TPA) for being 20 mmol/L by 0.5 mL concentration, 0.5 mL concentration are the ascorbic acid and 50 of 0.8 mmol/L
μ L concentration is that be added to the pH that 2.95 mL concentration are 200 mmol/L be 8.0 to nano cupric oxide made from 100 mg/L examples 1
Phosphate buffer in, mixing shakes up and is placed on 45 DEG C of warm bath, measures its fluorescence intensity at 421 nm after ten minutes
(excitation wavelength is 315 nm).It is repeated 6 times, the relative standard deviation of testing result is 0.54 %.
Example 11:
The terephthalic acid (TPA) for being 20 mmol/L by 0.5 mL concentration, 50 μ L concentration are nano oxygen made from 100 mg/L examples 1
Change copper and 0.5 mL different material (uric acid of 10 mmol/L, the citric acid of 10 mmol/L, the oxalic acid of 10 mmol/L, 10
The glutathione of mmol/L, the glycine of 10 mmol/L, the alanine of 10 mmol/L, 10 mmol/L ethyl alcohol, 100 mmol/L
Sodium chloride, the glucose of 100 mmol/L, 100 mmol/L maltose, the lactose of 100 mmol/L, 100 mmol/L fruit
The cysteine of sugar or 1 mmol/L) replacing ascorbic acid, to be added to the pH that 2.95 mL concentration are 200 mmol/L be 8.0
In phosphate buffer, mixing, which shakes up, is placed on 45 DEG C of warm bath, measures its fluorescence intensity at 421 nm after ten minutes and (swashs
Hair wavelength is 315 nm).As shown in figure 9, compared with the signal that the ascorbic acid of 0.8 mmol/L generates, above-mentioned interfering substance institute
The signal of generation is negligible.
Example 12:
The terephthalic acid (TPA) for being 20 mmol/L by 0.5 mL concentration, 0.5 mL dilute 40 times of the drug containing ascorbic acid
Or orange juice solution, 50 μ L concentration are that be added to 2.95 mL concentration be 200 to nano cupric oxide made from 550 mg/L examples 1
In the phosphate buffer that the pH of mmol/L is 8.0, mixing, which shakes up, is placed on 45 DEG C of warm bath, measures it after ten minutes 421
Fluorescence intensity at nm (excitation wavelength is 315 nm).Through 9 gained ascorbic acid standard curve of embodiment be calculated drug or
The number that iodo- sodium thiosulfate back titration method in ascorbic acid content in orange juice, this numerical value and theoretical value and Chinese Pharmacopoeia obtains
According to consistent.Sample recovery rate 92.2%~100.0%, relative standard deviation 0.5-1.9%.It is examined by F and t is examined, two methods
It is 95% without marked difference in confidence level.It can be seen that this method is reliably applicable in.
Claims (10)
1. a kind of method for quickly measuring ascorbic acid with nano cupric oxide self-activation cascade catalytic fluorometry system, it is characterized in that anti-
Bad hematic acid, nano cupric oxide and terephthalic acid (TPA) three mix warm bath, its fluorescence exciting wavelength of reaction product and launch wavelength point
It Wei not 315 nm and 421 nm；Transmitting fluorescence intensity is detected through sepectrophotofluorometer, and nano cupric oxide can significantly increase fluorescence
Signal strength, intensification factor are about 41 times.
2. according to claim 1 quickly measure ascorbic acid with nano cupric oxide self-activation cascade catalytic fluorometry system
Method, it is characterized in that the pH value of reaction system is 8.0.
3. according to claim 1 quickly measure ascorbic acid with nano cupric oxide self-activation cascade catalytic fluorometry system
Method, it is characterized in that bath temperature is 45 DEG C in reaction system, the reaction time is 10 minutes.
4. according to claim 1 quickly measure ascorbic acid with nano cupric oxide self-activation cascade catalytic fluorometry system
Method, it is characterized in that the concentration of nano cupric oxide is 1.25 mg/L in reaction system.
5. according to claim 1 quickly measure ascorbic acid with nano cupric oxide self-activation cascade catalytic fluorometry system
Method, it is characterized in that the P-phthalic acid at concentration in reaction system is 2.5 mmol/L.
6. a kind of method for quickly measuring ascorbic acid with nano cupric oxide self-activation cascade catalytic fluorometry system, it is characterized in that
It is separately added into ascorbic acid solution, phosphate buffer, terephthalic acid (TPA), the nano cupric oxide of various concentration in EP pipe, mixes
Liquid warm bath, reaction product is put into sepectrophotofluorometer and detects emitted luminescence intensity.
7. according to claim 6 quickly measure ascorbic acid with nano cupric oxide self-activation cascade catalytic fluorometry system
Method, it is characterized in that the ascorbic acid solution volume for the various concentration being added is 0.5 ml, the phosphate-buffered liquid of addition
Product is 2.95 ml, and concentration is 200 mmol/L, and 8.0,20 mmol/L terephthalic acid (TPA) volume of pH is 0.5 ml and 100
The nano oxidized copper volume of mg/L is 50 μ l, and mixed liquor warm bath at 45 DEG C, product fluorescence intensity is through fluorescence point after ten minutes
Light photometric determination maps to obtain standard curve, fluorescence intensity and ascorbic acid concentrations with fluorescence intensity Ascorbic Acid concentration
7.5 × 10-7-7.5×10-6M and 1.25 × 10-5-1.25×10-4It is in a linear relationship within the scope of M, detection is limited to 2.92 ×
10-8 M。
8. a kind of method for quickly measuring ascorbic acid with nano cupric oxide self-activation cascade catalytic fluorometry system, it is characterized in that step
It is rapid as follows: (1) drug to be added to ultrasonic dissolution in distilled water, drug sample solution is obtained after dilution；Orange juice directly dilutes
Juice samples solution；(2) drug sample solution or juice samples solution, nano cupric oxide and terephthalic acid (TPA) three are mixed into temperature
Bath；(3) reaction product is placed in fluorescence intensity in sepectrophotofluorometer, medicine is calculated according to the linearity curve of ascorbic acid
Product or juice samples content.
9. according to claim 8 quickly measure ascorbic acid with nano cupric oxide self-activation cascade catalytic fluorometry system
Method, it is characterized in that: (1) drug is added to ultrasonic dissolution in 50 mL distilled water, it is molten to obtain drug sample for 40 times of dilution after standing
Liquid；Orange juice directly dilutes 40 times and obtains juice samples solution；(2) by 0.5 mL drug sample solution or juice samples solution, volume
The nano cupric oxide that for 0.5 ml concentration be 20 mmol/L terephthalic acid (TPA)s and 100 mg/L volumes are 50 μ l is added to volume
For 2.95 ml, concentration is 200 mmol/L, mixes and is placed in 45 DEG C of water-baths 10 minutes in the phosphate buffer that pH is 8.0；
(3) reaction product is placed in fluorescence intensity in sepectrophotofluorometer, drug is measured according to the linearity curve of ascorbic acid
Or ascorbic acid content in orange juice.
10. according to claim 8 or claim 9 with nano cupric oxide self-activation cascade catalytic fluorometry system quickly measure Vitamin C
The method of acid, it is characterized in that nano cupric oxide is specifically by following step preparation: (1) taking the acetic acid copper solution of 0.02 mol/L
150 ml and 0.5 ml glacial acetic acid are added in the three-necked bottle equipped with condenser pipe, are heated with stirring to boiling；(2) 0.04 is rapidly joined
10 ml of sodium hydroxide solution of g/ml continues stirring 5 minutes after adding, obtain brown copper oxide precipitating；(3) reaction is obtained
Cupric oxide precipitate centrifugation, wash three times with dehydrated alcohol, be dried under reduced pressure to get diameter for 6 nm nano oxidized copper powder
Body disperses nano-cupric oxide powder in secondary distilled water and obtains brown nano cupric oxide colloidal solution.
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