CN110907406A - Method for measuring trace hydrogen peroxide based on mimic enzyme - Google Patents
Method for measuring trace hydrogen peroxide based on mimic enzyme Download PDFInfo
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- CN110907406A CN110907406A CN201911175386.9A CN201911175386A CN110907406A CN 110907406 A CN110907406 A CN 110907406A CN 201911175386 A CN201911175386 A CN 201911175386A CN 110907406 A CN110907406 A CN 110907406A
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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
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/47—Scattering, i.e. diffuse reflection
- G01N21/49—Scattering, i.e. diffuse reflection within a body or fluid
- G01N21/51—Scattering, i.e. diffuse reflection within a body or fluid inside a container, e.g. in an ampoule
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/38—Diluting, dispersing or mixing samples
Abstract
The invention discloses a method for measuring trace hydrogen peroxide based on mimic enzyme, which is characterized in that B-R buffer solution and β -CD-FeC are respectively added into an 11-branch colorimetric tube4Pc mixture, OP solution and OPD solution, 0.0, 0.0001, 0.001, 0.005, 0.01, 0.02, 0.04, 0.08, 0.1, 0.12, 0.14mL 8.0X 10‑6mol/L H2O2The solution was reacted in a water bath for 20 minutes, and then the nanogold solution was added, diluted to 5mL after 10 minutes, and the scattering spectrum was scanned on a fluorescence photometer at λ ex = λ em. Respectively measuring the addition of H at 700nm2O2Scattering intensity of solution I700nmAnd without addition of H2O2Scattering intensity of reagent blank I0Calculating the difference DeltaI700nm=I700nm‑I0. The determination method has good selectivity and high sensitivity.
Description
Technical Field
The invention relates to a method for measuring trace hydrogen peroxide, in particular to a resonance scattering spectrometry method for measuring hydrogen peroxide by using a compound formed by β -cyclodextrin, polyethylene glycol octyl phenyl ether and tetracarboxyl iron phthalocyanine as a simulated enzyme.
Background
The hydrogen peroxide is used as a strong oxidant, has the functions of sterilization, disinfection, bleaching and the like, and is widely applied to the industries of medicine, food, chemical industry and the like. Hydrogen peroxide is ubiquitous in the atmosphere and rain and is a major cause of acid rain. Therefore, the method has important significance for detecting the hydrogen peroxide. Resonance scattering spectroscopy has been used for analysis of many substances because of its advantages such as simple operation, high sensitivity, and low detection limit, and some reports have been made on the measurement of hydrogen peroxide. Tetracarboxyferric phthalocyanine (FeC)4Pc) is a peroxide mimic enzyme which can catalyze hydrogen peroxide to oxidize o-phenylenediamine (OPD) to generate an intermediate product 2, 3-diamino-5, 10-dihydrophenazine and a final product 2, 3-diaminophenazine, wherein 2 products can induce nanogold to aggregate so as to enhance the resonance scattering strength of the system, β -cyclodextrin (β -CD) and polyethylene glycol octyl phenyl ether (OP) are added into the reaction system, and FeC is4Pc, β -CD and OP generate OP- β -CD-FeC4The Pc compound can change the microenvironment of the mimic enzyme in the catalytic reaction, and further improve the catalytic activity of the mimic enzyme. A new method for quantitatively determining the trace hydrogen peroxide can be established according to the change of the resonance scattering intensity of the system before and after the reaction, and the method is not reported at present.
Disclosure of Invention
The invention aims to provide a resonance scattering spectrometry for measuring hydrogen peroxide by taking a compound formed by β -cyclodextrin, polyethylene glycol octyl phenyl ether and tetracarboxyl iron phthalocyanine as a simulated enzyme.
The method comprises the following specific steps:
(1) mixing β -CD saturated solution with concentration of 1.0 × 10-6FeC in mol/L4Pc is mixed evenly according to the volume ratio of 1:3 and placed for 12 hours to obtain β -CD-FeC4Pc to mix the solution.
(2) To a 5mL cuvette of 11 cells, 0.06mL of a Bertany-Robinson (B-R) buffer solution having a pH of 5.0 and 0.12mL of β -CD-FeC obtained in step (1) were added4Pc mixed solution, 0.04mL of polyethylene glycol octyl phenyl ether (OP) solution with concentration of 0.5g/L and 0.9mL of 0.01Adding 0.0, 0.0001, 0.001, 0.005, 0.01, 0.02, 0.04, 0.08, 0.1, 0.12 and 0.14mL of o-phenylenediamine (OPD) solution in sequence at a concentration of 8.0 × 10-6mol/L of H2O2After the solution was reacted in a water bath at 27 ℃ for 20 minutes, 0.6mL of a 58.0 μ g/mL nanogold (AuNPs) solution was added to each cuvette, shaken, immediately diluted to a predetermined volume with redistilled water, and left at room temperature for 10 minutes, and then a resonance scattering spectrum was scanned with a 1cm quartz cuvette using a fluorescence spectrophotometer at λ ex ═ λ em. The addition of H is measured at a wavelength of 700nm2O2Resonance scattering intensity of solution I700nmAnd without addition of H2O2The resonance scattering intensity I of the reagent blank solution0Calculating the difference DeltaI of the resonance scattering intensity700nm＝I700nm-I0Difference of resonant scattering intensity Δ I700nmAnd H2O2The concentration c is 1.60X 10-10～2.24×10-7Linear relation in mol/L range, linear regression equation is that delta I is 0.8420 multiplied by 109c-0.6400, correlation 0.9983, detection limit 8.90X 10-11mol/L; filtering pond water, fresh rainwater and river water, measuring resonance scattering intensity value by the same method with 0.2mL of filtrate, and calculating H in pond water, fresh rainwater and river water2O2The content of (a).
The determination method has good selectivity and high sensitivity.
Drawings
FIG. 1 shows a blank and 6.4X 10 of an embodiment of the present invention-8mol/L H2O2The resonance scattering spectrogram of (1);
the label in the figure is a blank, b is pH5.0B-R buffer solution-0.5 g/L OP-0.01mol/L OPD-OP- β -CD-FeC4Pc-6.4×10-8mol/L H2O2-AuNPs。
Detailed Description
Example (b):
(1) mixing β -CD saturated solution with concentration of 1.0 × 10-6FeC in mol/L4Pc is mixed evenly according to the volume ratio of 1:3 and placed for 12 hours to obtain β -CD-FeC4Pc to mix the solution.
(2) To a 5mL cuvette of 11 cells, 0.06mL of a Bertany-Robinson (B-R) buffer solution having a pH of 5.0 and 0.12mL of β -CD-FeC obtained in step (1) were added4Pc mixed solution, 0.04mL OP solution with concentration of 0.5g/L and 0.9mL OPD solution with concentration of 0.01mol/L, then 0.0, 0.0001, 0.001, 0.005, 0.01, 0.02, 0.04, 0.08, 0.1, 0.12, 0.14mL OPD solution with concentration of 8.0 × 10-6mol/L H2O2After the solution was reacted in a water bath at 27 ℃ for 20 minutes, 0.6mL of a 58.0 μ g/mL nanogold (AuNPs) solution was added to each cuvette, shaken, immediately diluted to a predetermined volume with redistilled water, and left at room temperature for 10 minutes, and then a resonance scattering spectrum was scanned with a 1cm quartz cuvette using a fluorescence spectrophotometer at λ ex ═ λ em. The addition of H is measured at a wavelength of 700nm2O2Resonance scattering intensity of solution I700nmAnd without addition of H2O2The resonance scattering intensity I of the reagent blank solution0Calculating the difference DeltaI of the resonance scattering intensity700nm＝I700nm-I0Difference of resonant scattering intensity Δ I700nmAnd H2O2The concentration c is 1.60X 10-10～2.24×10- 7Linear relation in mol/L range, linear regression equation of △ I0.8420X 109c-0.6400, correlation 0.9983, detection limit 8.90X 10-11mol/L; filtering pond water, fresh rainwater and river water, measuring resonance scattering intensity value with the same method with 0.2mL of filtrate, substituting into the regression equation, and calculating H in pond water, fresh rainwater and river water2O2The content of (a). The results of the spiking recovery experiments are shown in Table 1.
TABLE 1 Water sample analysis results (n ═ 5)
| Sample (I) | Measured value (10)-8mol/L) | Recovery (%) | RSD(％) |
| Pond water | 3.93 | 98.03 | 2.1 |
| Rain water | 2.62 | 101.17 | 3.2 |
| River water | 2.35 | 100.33 | 3.2 |
。
Claims (1)
1. A method for measuring trace hydrogen peroxide is characterized by comprising the following specific steps:
(1) mixing β -CD saturated solution with concentration of 1.0 × 10-6FeC in mol/L4Pc is mixed evenly according to the volume ratio of 1:3 and placed for 12 hours to obtain β -CD-FeC4Mixing a Pc mixed solution;
(2) to 11 tubes of 5mL colorimetric tube were added 0.06mL of Bertanin-Robinson buffer solution with pH =5.0 and 0.12mL of β -CD-FeC obtained in step (1)4Pc mixed solution, 0.04mL of polyethylene glycol octyl phenyl ether solution with concentration of 0.5g/L and 0.9mL of o-phenylenediamine solution with concentration of 0.01mol/L, then 0.0, 0.0001, 0.001, 0.005, 0.01, 0.02, 0.04, 0.08, 0.1, 0.12 and 0.14mL of o-phenylenediamine solution with concentration of 8.0 × 10 are added in sequence-6mol/L of H2O2The solution is reacted in a water bath at 27 DEG CAfter 20 minutes, adding 0.6mL of 58.0 mu g/mL nanogold solution into each colorimetric tube, shaking up, immediately diluting to a scale with secondary distilled water, standing at room temperature for 10 minutes, and scanning a resonance scattering spectrum on a fluorescence photometer by using a 1cm quartz cuvette in a mode of lambda ex = lambda em; the addition of H is measured at a wavelength of 700nm2O2Resonance scattering intensity of solution I700nmAnd without addition of H2O2The resonance scattering intensity I of the reagent blank solution0Calculating the difference DeltaI of the resonance scattering intensity700nm=I700nm-I0Difference of resonant scattering intensity Δ I700nmAnd H2O2The concentration c is 1.60X 10-10～2.24×10-7Linear in mol/L range, and the linear regression equation is delta I =0.8420 multiplied by 109c-0.6400, correlation 0.9983, detection limit 8.90X 10-11mol/L; filtering pond water, fresh rainwater and river water, measuring resonance scattering intensity value by the same method with 0.2mL of filtrate, and calculating H in pond water, fresh rainwater and river water2O2The content of (a).
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Citations (4)
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| CN101450798A (en) * | 2007-11-29 | 2009-06-10 | 索尼株式会社 | Method for processing carbon nanotube, carbon nanotube and carbon nanotube element |
| CN101583425A (en) * | 2005-02-17 | 2009-11-18 | 孟山都技术公司 | Transition metal-containing catalysts and catalyst combinations including transition metal-containing catalysts and processes for their preparation and use as oxidation catalysts |
| CN108254344A (en) * | 2018-01-03 | 2018-07-06 | 桂林理工大学 | A kind of method that melamine is measured using crystal violet |
| CN108318456A (en) * | 2018-01-03 | 2018-07-24 | 桂林理工大学 | A method of roxithromycin is measured based on eosin |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101583425A (en) * | 2005-02-17 | 2009-11-18 | 孟山都技术公司 | Transition metal-containing catalysts and catalyst combinations including transition metal-containing catalysts and processes for their preparation and use as oxidation catalysts |
| CN101450798A (en) * | 2007-11-29 | 2009-06-10 | 索尼株式会社 | Method for processing carbon nanotube, carbon nanotube and carbon nanotube element |
| CN108254344A (en) * | 2018-01-03 | 2018-07-06 | 桂林理工大学 | A kind of method that melamine is measured using crystal violet |
| CN108318456A (en) * | 2018-01-03 | 2018-07-24 | 桂林理工大学 | A method of roxithromycin is measured based on eosin |
Non-Patent Citations (4)
| Title |
|---|
| AIHUI LIANG ET AL.，: ""A Sensitive Resonance Scattering Spectral Assay for the Determination of Trace H2O2 Based on the HRP Catalytic Reaction and Nanogold Aggregation"", 《JOURNAL OF FLUORESCENCE》 * |
| 夏海鸣: "四羧基铁酞菁作为过氧化物模拟酶光谱法测定过氧化氢", 《万方学位论文》 * |
| 庄绪杰 等: "β－环糊精／聚乙二醇复合物的制备及表征", 《化学世界》 * |
| 郑琦 等: "过氧化物模拟酶的研究—四磺基铁酞菁与β-CD 的包合物的分析应用", 《江汉大学学报》 * |
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