CN104990913B - A kind of method based on gold nano grain detection of the growth glucose - Google Patents
A kind of method based on gold nano grain detection of the growth glucose Download PDFInfo
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- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 title claims abstract description 65
- 239000008103 glucose Substances 0.000 title claims abstract description 65
- 238000001514 detection method Methods 0.000 title claims abstract description 26
- 238000000034 method Methods 0.000 title claims abstract description 23
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 title claims abstract description 14
- 239000010931 gold Substances 0.000 title claims abstract description 14
- 229910052737 gold Inorganic materials 0.000 title claims abstract description 14
- 239000002245 particle Substances 0.000 claims abstract description 28
- FDWREHZXQUYJFJ-UHFFFAOYSA-M gold monochloride Chemical compound [Cl-].[Au+] FDWREHZXQUYJFJ-UHFFFAOYSA-M 0.000 claims abstract description 22
- 230000012447 hatching Effects 0.000 claims abstract description 19
- 241001247197 Cephalocarida Species 0.000 claims abstract description 16
- 238000002835 absorbance Methods 0.000 claims abstract description 12
- 239000008363 phosphate buffer Substances 0.000 claims abstract description 9
- 238000011534 incubation Methods 0.000 claims abstract description 6
- 235000009754 Vitis X bourquina Nutrition 0.000 claims description 10
- 235000012333 Vitis X labruscana Nutrition 0.000 claims description 10
- 235000014787 Vitis vinifera Nutrition 0.000 claims description 10
- 101710128063 Carbohydrate oxidase Proteins 0.000 claims description 3
- SJUCACGNNJFHLB-UHFFFAOYSA-N O=C1N[ClH](=O)NC2=C1NC(=O)N2 Chemical compound O=C1N[ClH](=O)NC2=C1NC(=O)N2 SJUCACGNNJFHLB-UHFFFAOYSA-N 0.000 claims description 3
- 235000013339 cereals Nutrition 0.000 claims description 3
- 240000007594 Oryza sativa Species 0.000 claims description 2
- 235000007164 Oryza sativa Nutrition 0.000 claims description 2
- 239000007788 liquid Substances 0.000 claims description 2
- 235000009566 rice Nutrition 0.000 claims description 2
- 240000006365 Vitis vinifera Species 0.000 claims 2
- 108010015776 Glucose oxidase Proteins 0.000 abstract description 24
- 239000004366 Glucose oxidase Substances 0.000 abstract description 24
- 229940116332 glucose oxidase Drugs 0.000 abstract description 24
- 235000019420 glucose oxidase Nutrition 0.000 abstract description 24
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 abstract description 23
- 230000008859 change Effects 0.000 abstract description 8
- 102000004190 Enzymes Human genes 0.000 abstract description 6
- 108090000790 Enzymes Proteins 0.000 abstract description 6
- 229940088598 enzyme Drugs 0.000 abstract description 6
- 230000008569 process Effects 0.000 abstract description 4
- RGHNJXZEOKUKBD-UHFFFAOYSA-N D-gluconic acid Natural products OCC(O)C(O)C(O)C(O)C(O)=O RGHNJXZEOKUKBD-UHFFFAOYSA-N 0.000 abstract description 3
- RGHNJXZEOKUKBD-SQOUGZDYSA-N Gluconic acid Natural products OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C(O)=O RGHNJXZEOKUKBD-SQOUGZDYSA-N 0.000 abstract description 3
- 235000012208 gluconic acid Nutrition 0.000 abstract description 3
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- 230000008901 benefit Effects 0.000 abstract description 2
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- 229960002163 hydrogen peroxide Drugs 0.000 description 10
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- 239000008280 blood Substances 0.000 description 8
- 210000004369 blood Anatomy 0.000 description 8
- 206010012601 diabetes mellitus Diseases 0.000 description 8
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Abstract
A kind of method based on gold nano detection of the growth glucose, step are as follows：1) glucose oxidase and concentration are separately added into phosphate buffer for 0 1.0mM glucose solution, hatch 10min at room temperature, obtain artemia hatching solution；2) 5nm golden nanometer particles and gold chloride are added in artemia hatching solution, incubation at room temperature 30min, shoots photo and read their corresponding absorbances at 530nm with the microplate readers of Synergy 2.It is an advantage of the invention that：The detection method is catalyzed glucose by glucose oxidase and generates gluconic acid, produce hydrogen peroxide and reduce gold chloride and generate golden simple substance, 5nm golden nanometer particle is promoted to grow, color change is produced simultaneously, the measure of glucose content is carried out with this, a kind of enzyme of glucose oxidase is only needed in whole detection process, expensive instrument need not be used, not only reduce cost, and quickly, it is convenient, it is easy to implement.
Description
Technical field
The present invention relates to the method for quantitative detection glucose content, particularly one kind is based on gold nano detection of the growth glucose
Method.
Background technology
With the improvement of people ' s living standards, the reduction of change and the manual labor of dietary structure, the incidence of disease of diabetes
It is growing day by day.At present, the whole world there are about diabetic more than 200,000,000, it has also become one of major disease of human health is threatened, in addition,
Diabetes can also trigger a series of complication, such as hypertension, kidney failure, blindness, apoplexy, heart disease, and therefore, diabetes are examined
Disconnected and treatment always is the key subjects that world medicine faces.Blood sugar concentration is an important clinical indices of diabetes,
The early diagnosis of diabetes can realize that, in order to better control over this disease, diabetic needs by detecting blood glucose
Its blood-sugar content is determined daily.Therefore, side that is simple, cheap and being effectively used for monitoring glucose level in patient blood is developed
Method is all very important in family expenses detection and Clinical screening.
In recent years, to improve sensitivity, specificity and the reliability of glucose sensor, scientists have been paid a lot
Effort.In these sensing technologies, most representational is blood glucose meter, and it is played in monitoring diabetes and offer clinical information
Very important effect.However, blood glucose meter relies in electrochemistry, electrode is costly and is easily inactivated in detection process by matrix, leads
Cause measurement result inaccurate.In addition, blood glucose meter is mainly to be family expenses detection design, clinical laboratory is not particularly suited for great amount of samples
The high flux examination of progress.
At present, the most frequently used method of the Clinical screening of blood glucose is the assay method based on colorimetric.These methods all pass through
Detection glucose oxidase is catalyzed hydrogen peroxide caused by glucose to detect blood glucose.And the detection of hydrogen peroxide then needed
Its chromogen substrate of oxide enzymatic generates colored solutions in the presence of hydrogen peroxide.One drawback of this method is and Portugal
Grape carbohydrate oxidase is compared, and peroxidase is very expensive, and its is active affected by environment larger, inconvenient for use and cost compared with
It is high.
The content of the invention
The present invention seeks to overcome the shortcomings of the prior art, there is provided a kind of quick, convenient, cheap based on gold nano
Glucose sensing approach is grown, this method is catalyzed glucose by glucose oxidase and generates gluconic acid, while produces peroxide
Change hydrogen, hydrogen-peroxide reduction gold chloride generates golden simple substance, promotes the growth of golden nanometer particle, while produces color change, with this
To carry out the measure of glucose content.Only need to use a kind of enzyme of glucose oxidase in whole detection process, testing result can
Depending on change, it is not necessary to expensive instrument, reduce testing cost.
Technical scheme：
A kind of method based on gold nano detection of the growth glucose, comprises the following steps：
1) by the glucose solution of one group of certain concentration in concentration is the phosphate buffer that 0.01M, pH are 7.2-7.4
Mixed respectively with glucose oxidase (GOx), hatch 2-10min at room temperature, obtain one group of artemia hatching solution a；
2) 5nm golden nanometer particles and gold chloride are added separately in above-mentioned each artemia hatching solution, incubation at room temperature 5-60min, obtained
To one group of artemia hatching solution b, artemia hatching solution b photos are shot respectively and read their phases at 530nm with the microplate readers of Synergy 2
The absorbance answered, according to concentration of glucose and its corresponding absorbance fit standard curve；
3) by the glucose solution sample of concentration to be measured in concentration is the phosphate buffer that 0.01M, pH are 7.2-7.4
Mixed with glucose oxidase (GOx), hatch 2-10min at room temperature, add 5nm golden nanometer particle and chlorauric acid solution, room
Temperature hatching 5-60min, reads its corresponding absorbance at 530nm with the microplate readers of Synergy 2, utilizes above-mentioned mark
Directrix curve, calculate the concentration of glucose solution sample to be measured.
The concentration of glucose oxidase is 1.0-3.0mg/mL in the step 1) artemia hatching solution；The glucose of certain concentration is molten
The concentration of liquid is respectively 0,0.1,0.3,0.5,0.7,0.9,1.0mM.
The concentration of 5nm golden nanometer particles is 4-10nM in step 2) the artemia hatching solution b, and the concentration of gold chloride is 0.05-
0.4mM。
The concentration of golden nanometer particle and the concentration of gold chloride are identical with the concentration in step 2) in the step 3).
A kind of method based on gold nano detection of the growth glucose is as follows for heterogeneous experiment, step：
1) filter paper is broken into a diameter of 1cm filter paper with card punch；
2) in concentration is the phosphate buffer that 0.01M, pH are 7.3 by glucose oxidase (GOx), gold chloride and
5nm solution of gold nanoparticles mixing, obtains mixed liquor, mixed liquor is added dropwise on above-mentioned filter paper, be incubated at room temperature 2-
10min；
3) glucose solution of one group of certain concentration is added dropwise on the above-mentioned scraps of paper, is incubated at room temperature 5-60min, grape is glycoxidative
Enzymatic glucose produces hydrogen peroxide, hydrogen-peroxide reduction gold chloride, promotes golden nanometer particle to grow, filter paper gradually becomes
Red, the red weights of the scraps of paper and glucose concentration are into positive correlation.
4) mixed liquor in step 2) is added dropwise on filter paper, is incubated at room temperature 2-10min, the grape of concentration to be measured is added dropwise
Sugar juice, 5-60min is incubated at room temperature, color contrast is carried out with gained filter paper in step 3), qualitative detection glucose solution
Concentration.
The concentration of glucose oxidase is 1.0-3.0mg/mL in the step 2) mixed liquor, the end of 5nm golden nanometer particles
Concentration is 4-10nM, the final concentration of 0.05-0.4mM of gold chloride.
In the step 3) concentration of the glucose solution of certain concentration be respectively 0,1,10,20,50,100mM.
The working mechanism of the present invention：
Golden nanometer particle because it has very high extinction coefficient, be often used to build colorimetric methods come detect enzyme,
The multiple analytes such as DNA, organic molecule, ion.And the extinction coefficient of golden nanometer particle is closely related with its size, size is got over
Greatly, extinction coefficient is bigger, and therefore, at low concentrations, small size collaurum is colourless, and the large scale colloid under same concentrations
Jin Ze is red.Using hydrogen-peroxide reduction gold chloride caused by glucose oxidase catalysis glucose in the present invention, promote
The growth of gold nano grain, so as to produce color change to carry out the measure of glucose content.
It is an advantage of the invention that：
The detection method is catalyzed glucose by glucose oxidase and generates gluconic acid, while produces hydrogen peroxide, mistake
Hydrogen oxide can reduce gold chloride and generate golden simple substance, promote 5nm golden nanometer particle to grow, while produce color change, with this
To carry out the measure of glucose content.Only need to use a kind of enzyme of glucose oxidase in whole detection process, testing result can
Depending on change, it is not necessary to expensive instrument, not only reduce testing cost, and quickly, it is convenient, it is easy to implement.
Brief description of the drawings
Fig. 1 is the method schematic diagram of the detection of the growth glucose based on golden nanometer particle.
Fig. 2 be concentration be 0,0.1,0.3,0.5,0.7,0.9, the testing result colour developing figure of 1.0mM glucose solution.
Fig. 3 is the glucose solution of various concentrations in A530nmThe fitting a straight line figure at place, wherein A530nmIt is glycoxidative to grape
After gold chloride and 5nm golden nanometer particles room temperature reaction 30min are added in enzyme and glucose mixed solution micropore is used at 530nm
Plate reader reads absorbance.
Fig. 4 is result colour developing figure of the method for the detection of the growth glucose based on golden nanometer particle on filter paper.
Embodiment
The present invention is further described with reference to embodiment.Unless separately have other explanations in the application context,
Technical term use herein and abbreviation are respectively provided with conventional sense known to those skilled in the art；Unless otherwise indicated, under
It is commercially available to state raw materials used compound in embodiment.
Embodiment：
A kind of method based on gold nano detection of the growth glucose, comprises the following steps：
1) glucose solution of one group of certain concentration is distinguished in concentration is the phosphate buffer that 0.01M, pH are 7.4
Mixed with glucose oxidase (GOx), hatch 10min at room temperature, obtain one group of artemia hatching solution a, the glucose solution of certain concentration
Concentration be respectively 0,0.1,0.3,0.5,0.7,0.9,1.0mM；
2) 5nm golden nanometer particles and gold chloride are added separately in above-mentioned each artemia hatching solution, 5nm gold nanos in artemia hatching solution b
The concentration of particle is 8.3nM, and the concentration of gold chloride is 0.16mM, incubation at room temperature 30min, obtains one group of artemia hatching solution b, shoots respectively
Artemia hatching solution b photos simultaneously read their corresponding absorbances at 530nm with the microplate readers of Synergy 2, according to grape
Sugared concentration and its corresponding absorbance fit standard curve；
3) by the glucose solution sample of concentration to be measured in concentration is the phosphate buffer that 0.01M, pH are 7.2-7.4
Mixed with glucose oxidase (GOx), hatch 10min at room temperature, add 5nm golden nanometer particle and chlorauric acid solution, Jenner
The concentration of rice corpuscles and the concentration of gold chloride are identical with the concentration in step 2), incubation at room temperature 30min, with the micropores of Synergy 2
Plate reader reads its corresponding absorbance at 530nm, using above-mentioned standard curve, calculates glucose solution sample to be measured
The concentration of product.
Fig. 1 is the method schematic diagram of the detection of the growth glucose based on golden nanometer particle.
Fig. 2 be concentration be 0,0.1,0.3,0.5,0.7,0.9, the testing result colour developing figure of 1.0mM glucose solution, figure
In show：With the increase of concentration of glucose, golden nanometer particle is gradually grown up, and solution colour gradually reddens.
Fig. 3 is the glucose solution of various concentrations in A530nmThe fitting a straight line figure at place, wherein A530nmIt is glycoxidative to grape
After gold chloride and 5nm golden nanometer particles room temperature reaction 30min are added in enzyme and glucose mixed solution micropore is used at 530nm
Plate reader reads absorbance.Show in figure：After growing 30min, A530nmIt is linear with the concentration of glucose.
A kind of method based on gold nano detection of the growth glucose is as follows for heterogeneous experiment, step：
1) filter paper is broken into a diameter of 1cm filter paper with card punch；
2) by glucose oxidase (GOx), gold chloride in concentration is the phosphate buffer that 0.01M, pH are 7.2-7.4
Mixed with 5nm solution of gold nanoparticles, obtain mixed liquor, the concentration of glucose oxidase is 1.0mg/mL, 5nm in mixed liquor
The final concentration of 8.3nM of golden nanometer particle, the final concentration of 0.16mM of gold chloride, mixed liquor is added dropwise on above-mentioned filter paper,
It is incubated at room temperature 10min；
3) glucose solution of one group of certain concentration is added dropwise on the above-mentioned scraps of paper, is incubated at room temperature 5-60min, grape is glycoxidative
Enzymatic glucose produces hydrogen peroxide, hydrogen-peroxide reduction gold chloride, promotes golden nanometer particle to grow, filter paper gradually becomes
Red, the red weights of the scraps of paper and glucose concentration are into positive correlation.
4) mixed liquor in step 2) is added dropwise on filter paper, is incubated at room temperature 2-10min, the grape of concentration to be measured is added dropwise
Sugar juice, 5-60min is incubated at room temperature, color contrast is carried out with gained filter paper in step 3), qualitative detection glucose solution
Concentration.
Fig. 4 is result colour developing figure of the method on filter paper of the detection of the growth glucose based on golden nanometer particle, table in figure
It is bright：With the increase of concentration of glucose, golden nanometer particle is gradually grown up, and the scraps of paper gradually redden.
Claims (1)
- A kind of 1. method based on gold nano detection of the growth glucose, it is characterised in that it is by following steps：1) in concentration is the phosphate buffer that 0.01M, pH are 7.4 by the glucose solution of one group of certain concentration respectively with Portugal Grape carbohydrate oxidase (GOx) mixes, and hatches 10min at room temperature, obtains one group of artemia hatching solution a, the glucose solution of certain concentration it is dense Degree respectively 0,0.1,0.3,0.5,0.7,0.9,1.0mM；2) 5nm golden nanometer particles and gold chloride are added separately in above-mentioned each artemia hatching solution, incubation at room temperature 30min, obtain one group The concentration of 5nm golden nanometer particles is 8.3nM in artemia hatching solution b, artemia hatching solution b, and the concentration of gold chloride is 0.16mM, respectively shooting hatching Liquid b photos simultaneously read their corresponding absorbances at 530nm with the microplate readers of Synergy 2, dense according to glucose Degree and its corresponding absorbance fit standard curve；3) by the glucose solution sample of concentration to be measured and Portugal in concentration is the phosphate buffer that 0.01M, pH are 7.2-7.4 Grape carbohydrate oxidase (GOx) mixes, and hatches 10min at room temperature, adds 5nm golden nanometer particle and chlorauric acid solution, Jenner's grain of rice The concentration of son and the concentration of gold chloride are identical with the concentration in step 2), incubation at room temperature 30min, are read with the microwell plates of Synergy 2 Number device reads its corresponding absorbance at 530nm, using above-mentioned standard curve, calculates glucose solution sample to be measured Concentration.
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| CN105296595B (en) * | 2015-10-29 | 2019-04-09 | 北京物资学院 | A kind of bioenzyme activity detection method based on nanogold growth |
| CN107561074A (en) * | 2017-09-27 | 2018-01-09 | 安徽师范大学 | A kind of biological colorimetric sensor, preparation method and its application for detecting glucose |
| GB2594619B (en) * | 2018-11-27 | 2022-12-07 | Module Innovations Private Ltd | Method for testing of antibiotic susceptibility in microorganisms |
| CN110672544A (en) * | 2019-10-12 | 2020-01-10 | 天津科技大学 | Glucose visualization sensor based on cyclic peptide recognition element and preparation method and application thereof |
| CN114295819B (en) * | 2021-12-30 | 2023-05-30 | 南京邮电大学 | Non-marking high-flux quantitative glucose detection method |
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| CN1912622A (en) * | 2006-08-10 | 2007-02-14 | 福建省洪诚生物药业有限公司 | Chemiluminescence investigating method of glucose in body fluid |
| CN103439482A (en) * | 2013-08-15 | 2013-12-11 | 中南大学 | Application of biosensing test paper based on N, N'-bis (trimethoxy silicyl propyl)-glutarimide |
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|---|---|---|---|---|
| CN1912622A (en) * | 2006-08-10 | 2007-02-14 | 福建省洪诚生物药业有限公司 | Chemiluminescence investigating method of glucose in body fluid |
| CN103439482A (en) * | 2013-08-15 | 2013-12-11 | 中南大学 | Application of biosensing test paper based on N, N'-bis (trimethoxy silicyl propyl)-glutarimide |
Non-Patent Citations (1)
| Title |
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| Glucose Oxidase-Catalyzed Growth of Gold Nanoparticles Enables Quantitative Detection of Attomolar Cancer Biomarkers;Dingbin Liu,et al;《analytical chemistry》;20140526;第5800页"Abstract"，5801页左栏第1段，第5801-5802页第"EXPERIMENTAL SECTION"节，第5802-5804页第"RESULTS AND DISCUSSION"节，Scheme 1，图1-图3 * |
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