CN105136784A - Preparation method and application of saccharide sensor based on colorimetric analysis - Google Patents
Preparation method and application of saccharide sensor based on colorimetric analysis Download PDFInfo
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- CN105136784A CN105136784A CN201510442852.0A CN201510442852A CN105136784A CN 105136784 A CN105136784 A CN 105136784A CN 201510442852 A CN201510442852 A CN 201510442852A CN 105136784 A CN105136784 A CN 105136784A
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Abstract
The invention relates to a preparation method and an application of a saccharide sensor based on colorimetric analysis and belongs to the technical fields of novel functional materials and novel sensor construction. In the invention, on the basis of quick response of a color developing composition on hydrogen peroxide, the saccharide sensor is significantly increased in sensitivity and reduced in detection limit. The sensor achieves real-time on-line detection on various saccharides and has important significance in detection of the saccharides and combined diagnosis with other marker substances.
Description
Technical field
The present invention relates to a kind of preparation method and application of the saccharide sensor device based on colorimetric analysis.Specifically adopt self-control ITO sensing platform to construct the colorimetric sensor detecting carbohydrate, belong to novel saccharide sensor detection technique field.
Background technology
Glucose is the index of blood-sugar content.In the process that blood sugar for human body content rises, along with the generation of some disease of human body, may usually show hypertension, high fat of blood, eye illness etc.Therefore, in clinical research, a kind of method developing quick, easy, sensitive detection glucose is very important.
Colorimetric analysis immunosensor has highly sensitive, Visual retrieval, easy and simple to handle, miniaturization, quick and precisely detects the advantages such as analysis, therefore the present invention has prepared a kind of colorimetric sensor based on colour developing compound, achieves the detection to dextrose plus saccharose.
The clinical testing procedure of current existing carbohydrate is a lot, as chemical analysis, silver mirror analysis, stratographic analysis etc.Colorimetric sensing analysis is a kind of sensor analytics method combined with visual colorimetric method, is combined by the catalysis characteristics between carbohydrate with enzyme, makes it have high sensitivity, high selectivity, analysis fast and the advantage such as Real-Time Monitoring.
The present invention adopts colour developing compound APTMS-GA to be base material, utilizes enzyme catalysis to build colorimetric sensor, produces hydrogen peroxide and colour developing compound is faded, achieve the Sensitive Detection of glucose by glucose oxidase to the catalysis of glucose.The present invention, also by the dual catalytic effect of enzyme, by glucose oxidase and invertase to the dual catalytic effect of sucrose, achieves the detection to sucrose.This approach enhance the sensitivity of sensor, widened the range of linearity, significantly reduce the detection limit of sensor, and can realize facilitating detection whenever and wherever possible.The method have highly sensitive, specificity good, detect the advantages such as quick, and achieves examinations, and do not need by large-scale instrument, effectively reduce making and the testing cost of sensor, effectively overcome the deficiency of current carbohydrate detection method.
Summary of the invention
An object of the present invention is base material based on colour developing compound APTMS-GA, construct a kind of can the colorimetric saccharide sensor device of Real-Time Monitoring.
Two of object of the present invention is exactly achieve the glucose detection on ITO sheet, achieves the highly sensitive detection of glucose.
Three of object of the present invention is exactly by glucose oxidase and the oxidasic dual catalytic effect of sucrose, achieves the highly sensitive detection to sucrose.
This colorimetric saccharide sensor device can be used for detecting in real time anywhere or anytime by four of object of the present invention exactly, reaches the effect of sensitivity monitoring.
technical scheme of the present invention is as follows:
1., based on a preparation method for the saccharide sensor device of colorimetric analysis, it is characterized in that, comprise the following steps:
(1) with massfraction be 0.1% ~ 10% the methanol solution of 3-aminopropyl triethoxysilane APTES soak the ito glass 24h of 3mm × 3mm, washing after taking out;
(2) on ito glass, drip 6 μ L, 3-aminopropyl trimethoxysilane-glutaraldehyde colour developing compound APTMS-GA, dry in 4oC refrigerator;
(3) on ITO, drip the 1-butyl-3-methyl imidazolium tetrafluoroborate BMIMBF of 1 ~ 20mg/mL
45% chitosan solution CS, form composite film IL-CS, dry in 4oC refrigerator;
(4) drip the golden nanometer particle of 6 μ L, dry in 4oC refrigerator;
(5) drip 6 μ L, different types of enzyme solutions, fully react, ultrapure water, the ITO modified is put into microwell plate successively, dry in 4oC refrigerator, obtained colorimetric saccharide sensor device.
2. the preparation method of a kind of saccharide sensor device based on colorimetric analysis as claimed in claim 1, described colour developing compound APTMS-GA, it is characterized in that, preparation process is as follows:
By massfraction be 0.5% ~ 12% 3-aminopropyl trimethoxysilane APTMS and massfraction be 0.2% ~ 10% glutaraldehyde GA mix according to 1:1 ratio, obtained bolarious APTMS-GA label.
3. the preparation method of a kind of saccharide sensor device based on colorimetric analysis as claimed in claim 1, described variety classes enzyme solutions, it is characterized in that, preparation process is as follows:
(1) preparation of glucose sensor
At the golden nanometer particle of dropping 6 μ L, after drying in 4oC refrigerator, continue the glucose oxidase solution of the 0.01-10mg/mL of dropping 6 μ L, dry in 4oC refrigerator; Continue the glucose solution of the 1-35mmol/L of dropping 6 μ L, obtained glucose colorimetric sensor;
(2) preparation of sucrose sensor
At the golden nanometer particle of dropping 6 μ L, after drying in 4oC refrigerator, continue the glucose oxidase solution of 0.01-10mg/mL and the 0.02-14mg/mL invertase solution of 6 μ L of dropping 6 μ L, dry in 4oC refrigerator; Continue the sucrose solution of the 1-50mmol/L of dropping 6 μ L, obtained sucrose in water ratio colour sensor.
4. the preparation method of a kind of saccharide sensor device based on colorimetric analysis as claimed in claim 1, for the detection of dextrose plus saccharose, detecting step is as follows:
After the ITO colour plate in microwell plate at room temperature fully reacts, under identical intensity of illumination, each block ITO is taken pictures, calculate the gray scale of each block glass with ImageJ software, obtain average gray and concentration of saccharide curve; According to the method for working curve, sample is measured, calculate the content of dextrose plus saccharose according to working curve.
useful achievement of the present invention
(1) the compound APTMS-GA that develops the color is good developer, very sensitive to the response of hydrogen peroxide, by glucose oxidase glucose output hydrogen peroxide, thus colour developing compound APTMS-GA is faded, carries out the quantitative detection to glucose.
(2) the present invention utilizes glucose oxidase and the oxidasic dual catalytic effect of sucrose first, achieves the Sensitive Detection to sucrose.
(3) use of ITO, instead of traditional electrode modification sensor, reach the convenient effect detected, and ITO is cheap, and single use, make sensor have good reappearance.
(4) the present invention utilizes the specific catalytic of enzyme to react, and improves the specificity of detection method.
(5) the present invention utilizes simple and easy photographing device, and contrast colors sensing chip is taken pictures, and carries out gray scale matching, realize Sensitive Detection by ImageJ software.Equipment is simple, and operation fast, is applicable to the prevention in advance that disease is carried out in the bad rural area of medical condition.
(6) the colorimetric immunoassay sensor prepared of the present invention is for the detection of dextrose plus saccharose, and the response time is short, and detectability is low, the range of linearity is wide, can realize simple, quick, highly sensitive and specific detection, the detectability of glucose reaches 0.3mmol/L, and the detectability of sucrose reaches 0.5mmol/L.
Embodiment
embodiment 1
A kind of preparation method of the saccharide sensor device based on colorimetric analysis and application
(1) with massfraction be 0.1% the methanol solution of 3-aminopropyl triethoxysilane APTES soak the ito glass 24h of 3mm × 3mm, washing after taking out;
(2) on ito glass, drip 6 μ L, 3-aminopropyl trimethoxysilane-glutaraldehyde colour developing compound APTMS-GA, dry in 4oC refrigerator;
(3) on ITO, drip the 1-butyl-3-methyl imidazolium tetrafluoroborate BMIMBF of 1mg/mL
45% chitosan solution CS, form composite film IL-CS, dry in 4oC refrigerator;
(4) drip the golden nanometer particle of 6 μ L, dry in 4oC refrigerator;
(5) drip 6 μ L, different types of enzyme solutions, fully react, ultrapure water, the ITO modified is put into microwell plate successively, dry in 4oC refrigerator, obtained colorimetric saccharide sensor device.
embodiment 2
A kind of preparation method of the saccharide sensor device based on colorimetric analysis and application
(1) with massfraction be 3% the methanol solution of 3-aminopropyl triethoxysilane APTES soak the ito glass 24h of 3mm × 3mm, washing after taking out;
(2) on ito glass, drip 6 μ L, 3-aminopropyl trimethoxysilane-glutaraldehyde colour developing compound APTMS-GA, dry in 4oC refrigerator;
(3) on ITO, drip the 1-butyl-3-methyl imidazolium tetrafluoroborate BMIMBF of 10mg/mL
45% chitosan solution CS, form composite film IL-CS, dry in 4oC refrigerator;
(4) drip the golden nanometer particle of 6 μ L, dry in 4oC refrigerator;
(5) drip 6 μ L, different types of enzyme solutions, fully react, ultrapure water, the ITO modified is put into microwell plate successively, dry in 4oC refrigerator, obtained colorimetric saccharide sensor device.
embodiment 3
A kind of preparation method of the saccharide sensor device based on colorimetric analysis and application
(1) with massfraction be 10% the methanol solution of 3-aminopropyl triethoxysilane APTES soak the ito glass 24h of 3mm × 3mm, washing after taking out;
(2) on ito glass, drip 6 μ L, 3-aminopropyl trimethoxysilane-glutaraldehyde colour developing compound APTMS-GA, dry in 4oC refrigerator;
(3) on ITO, drip the 1-butyl-3-methyl imidazolium tetrafluoroborate BMIMBF of 20mg/mL
45% chitosan solution CS, form composite film IL-CS, dry in 4oC refrigerator;
(4) drip the golden nanometer particle of 6 μ L, dry in 4oC refrigerator;
(5) drip 6 μ L, different types of enzyme solutions, fully react, ultrapure water, the ITO modified is put into microwell plate successively, dry in 4oC refrigerator, obtained colorimetric saccharide sensor device.
embodiment 4
A kind of preparation method of the saccharide sensor device based on colorimetric analysis and application
By massfraction be 0.5% 3-aminopropyl trimethoxysilane APTMS and massfraction be 0.2% glutaraldehyde GA mix according to 1:1 ratio, obtained bolarious APTMS-GA label.
embodiment 5
A kind of preparation method of the saccharide sensor device based on colorimetric analysis and application
By massfraction be 6% 3-aminopropyl trimethoxysilane APTMS and massfraction be 5% glutaraldehyde GA mix according to 1:1 ratio, obtained bolarious APTMS-GA label.
embodiment 6
A kind of preparation method of the saccharide sensor device based on colorimetric analysis and application
By massfraction be 12% 3-aminopropyl trimethoxysilane APTMS and massfraction be 10% glutaraldehyde GA mix according to 1:1 ratio, obtained bolarious APTMS-GA label.
embodiment 7
A kind of preparation method of the saccharide sensor device based on colorimetric analysis and application
(1) preparation of glucose sensor
At the golden nanometer particle of dropping 6 μ L, after drying in 4oC refrigerator, continue the glucose oxidase solution of the 0.01mg/mL of dropping 6 μ L, dry in 4oC refrigerator; Continue the glucose solution of the 1-35mmol/L of dropping 6 μ L, obtained glucose colorimetric sensor;
(2) preparation of sucrose sensor
At the golden nanometer particle of dropping 6 μ L, after drying in 4oC refrigerator, continue the glucose oxidase solution of 0.01mg/mL and the 0.02mg/mL invertase solution of 6 μ L of dropping 6 μ L, dry in 4oC refrigerator; Continue the sucrose solution of the 1-50mmol/L of dropping 6 μ L, obtained sucrose in water ratio colour sensor.
embodiment 8
A kind of preparation method of the saccharide sensor device based on colorimetric analysis and application
(1) preparation of glucose sensor
At the golden nanometer particle of dropping 6 μ L, after drying in 4oC refrigerator, continue the glucose oxidase solution of the 5mg/mL of dropping 6 μ L, dry in 4oC refrigerator; Continue the glucose solution of the 1-35mmol/L of dropping 6 μ L, obtained glucose colorimetric sensor;
(2) preparation of sucrose sensor
At the golden nanometer particle of dropping 6 μ L, after drying in 4oC refrigerator, continue the glucose oxidase solution of 5mg/mL and the 2mg/mL invertase solution of 6 μ L of dropping 6 μ L, dry in 4oC refrigerator; Continue the sucrose solution of the 1-50mmol/L of dropping 6 μ L, obtained sucrose in water ratio colour sensor.
embodiment 9
A kind of preparation method of the saccharide sensor device based on colorimetric analysis and application
(1) preparation of glucose sensor
At the golden nanometer particle of dropping 6 μ L, after drying in 4oC refrigerator, continue the glucose oxidase solution of the 10mg/mL of dropping 6 μ L, dry in 4oC refrigerator; Continue the glucose solution of the 1-35mmol/L of dropping 6 μ L, obtained glucose colorimetric sensor;
(2) preparation of sucrose sensor
At the golden nanometer particle of dropping 6 μ L, after drying in 4oC refrigerator, continue the glucose oxidase solution of 10mg/mL and the 14mg/mL invertase solution of 6 μ L of dropping 6 μ L, dry in 4oC refrigerator; Continue the sucrose solution of the 1-50mmol/L of dropping 6 μ L, obtained sucrose in water ratio colour sensor.
embodiment 10
A kind of preparation method of the saccharide sensor device based on colorimetric analysis and application
After the ITO colour plate in microwell plate at room temperature fully reacts, under identical intensity of illumination, each block ITO is taken pictures, calculate the gray scale of each block glass with ImageJ software, obtain average gray and concentration of saccharide curve; According to the method for working curve, sample is measured, calculate the content of dextrose plus saccharose according to working curve.
Claims (4)
1., based on a preparation method for the saccharide sensor device of colorimetric analysis, it is characterized in that, comprise the following steps:
(1) with massfraction be 0.1% ~ 10% the methanol solution of 3-aminopropyl triethoxysilane APTES soak the ito glass 24h of 3mm × 3mm, washing after taking out;
(2) on ito glass, drip 6 μ L, 3-aminopropyl trimethoxysilane-glutaraldehyde colour developing compound APTMS-GA, dry in 4oC refrigerator;
(3) on ITO, drip the 1-butyl-3-methyl imidazolium tetrafluoroborate BMIMBF of 1 ~ 20mg/mL
45% chitosan solution CS, form composite film IL-CS, dry in 4oC refrigerator;
(4) drip the golden nanometer particle of 6 μ L, dry in 4oC refrigerator;
(5) drip 6 μ L, different types of enzyme solutions, fully react, ultrapure water, the ITO modified is put into microwell plate successively, dry in 4oC refrigerator, obtained colorimetric saccharide sensor device.
2. the preparation method of a kind of saccharide sensor device based on colorimetric analysis as claimed in claim 1, described colour developing compound APTMS-GA, it is characterized in that, preparation process is as follows:
By massfraction be 0.5% ~ 12% 3-aminopropyl trimethoxysilane APTMS and massfraction be 0.2% ~ 10% glutaraldehyde GA mix according to 1:1 ratio, obtained bolarious APTMS-GA label.
3. the preparation method of a kind of saccharide sensor device based on colorimetric analysis as claimed in claim 1, described variety classes enzyme solutions, it is characterized in that, preparation process is as follows:
(1) preparation of glucose sensor
At the golden nanometer particle of dropping 6 μ L, after drying in 4oC refrigerator, continue the glucose oxidase solution of the 0.01-10mg/mL of dropping 6 μ L, dry in 4oC refrigerator; Continue the glucose solution of the 1-35mmol/L of dropping 6 μ L, obtained glucose colorimetric sensor;
(2) preparation of sucrose sensor
At the golden nanometer particle of dropping 6 μ L, after drying in 4oC refrigerator, continue the glucose oxidase solution of 0.01-10mg/mL and the 0.02-14mg/mL invertase solution of 6 μ L of dropping 6 μ L, dry in 4oC refrigerator; Continue the sucrose solution of the 1-50mmol/L of dropping 6 μ L, obtained sucrose in water ratio colour sensor.
4. the preparation method of a kind of saccharide sensor device based on colorimetric analysis as claimed in claim 1, for the detection of dextrose plus saccharose, detecting step is as follows:
After the ITO colour plate in microwell plate at room temperature fully reacts, under identical intensity of illumination, each block ITO is taken pictures, calculate the gray scale of each block glass with ImageJ software, obtain average gray and concentration of saccharide curve; According to the method for working curve, sample is measured, calculate the content of dextrose plus saccharose according to working curve.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108680569A (en) * | 2018-04-11 | 2018-10-19 | 天津市宝坻区人民医院 | Sucrose-determination method in serum |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108680569A (en) * | 2018-04-11 | 2018-10-19 | 天津市宝坻区人民医院 | Sucrose-determination method in serum |
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