CN106442674A - Electrochemical glucose biosensor based on strontium titanate nano particles and preparation method of biosensor - Google Patents
Electrochemical glucose biosensor based on strontium titanate nano particles and preparation method of biosensor Download PDFInfo
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Abstract
The invention relates to an electrochemical glucose biosensor and a preparation method of the biosensor. The preparation method of the electrochemical glucose biosensor includes the steps that firstly, strontium titanate nano particles are dispersed in a chitosan solution, and a GOD/SrTiO3/CS mixed solution is prepared; secondly, the prepared GOD/SrTiO3/CS mixed solution is modified to the surface of a glassy carbon electrode obtained after pretreatment, and the electrochemical glucose biosensor is obtained. The biosensor is easy and quick to prepare, low in cost, high in sensitivity and good in stability, the strontium titanate nano particles are used for fixing glucose oxidase, and biological catalytic activity of biomacromolecules can be well maintained.
Description
Technical field
The invention belongs to electrochemica biological sensor field, more particularly, to a kind of electrochemistry based on strontium titanates nano-particle
Glucose biological sensor and preparation method thereof.
Background technology
The health of the mankind in diabetes serious harm, and the therefore diagnosis for diabetes is particularly important.By detection
In blood of human body, the concentration of glucose can learn the physical condition of people indirectly, and the diagnosis for diabetes provides one to put down
Platform.
At present, the method for detection concentration of glucose has chromatography, AAS, Optical Rotation, colorimetric method etc., these points
Analysis method sensitivity is low, laborious, time-consuming, and has higher sensitivity using electrochemica biological sensor method, and analysis speed
Degree is fast, selective height, instrumentation are simple, lower-price characteristic.In recent years, electrochemical methods due to its sensitivity high,
Specificity is good, sample consumption is little, simple and quick the features such as, become the extremely competitive glucose in-situ check and test method of one kind.
Nano material has the features such as quantum size effect, surface-interface effect, bulk effect, has been used for preparing
Biology sensor.In recent years, metal nanoparticle, carbon nanomaterial and polymer nano material, have had been reported for electrification
Learn the preparation of glucose biological sensor.Although these materials have good performance, it is loaded down with trivial details to there is synthetic method, cost
Height, is difficult to be prepared on a large scale etc. not enough.Strontium titanates (SrTiO3) it is a kind of new semiconductor nano material, the former material of its preparation
Material aboundresources, with low cost, have a safety feature, pollution-free, the advantages of prepare easy.Additionally, it also has big specific surface
Long-pending, good biocompatibility and electric conductivity, can be used for fixing protein molecules, prepare biology sensor.At present, strontium titanates
Also no report is not used for fixing glucose oxidase, prepares electrochemica biological sensor.
Content of the invention
It is an object of the invention to propose a kind of electrochemical glucose biology sensor based on strontium titanates nano-particle and
Its preparation method.Strontium titanates nano-particle and glucose oxidase are modified on glass-carbon electrode, prepared electrochemical biosensor
Device；
The technical solution adopted in the present invention is：The invention provides a kind of electrochemistry Portugal based on strontium titanates nano-particle
Grape sugar biology sensor, is that jointly strontium titanates nano-particle and glucose oxidase are modified glass-carbon electrode, prepared electrochemistry
Biology sensor.
The invention also discloses the preparation method of the electrochemical glucose biology sensor of described strontium titanates nano-particle,
Comprise the following steps：
1) strontium titanates nano-particle is dispersed in chitosan solution, adds glucose oxidase, be configured to Portugal
Grape carbohydrate oxidase/strontium titanates nano-particle/shitosan mixed solution；
2) by step 1) in be obtained glucose oxidase/strontium titanates nano-particle/shitosan mixed solution uniformly coat
To the glassy carbon electrode surface through pretreatment, it is placed under 4 DEG C of temperature environment, after being dried, obtain described electrochemical glucose life
Thing sensor.
Described step 1) in, the preparation process of strontium titanates nano-particle is as follows：A) weigh the titanium dioxide of the amount of certain material
Strontium, the titanium dioxide (P25) of the amount of certain material and equimolar sodium chloride and potassium chloride, mix in mortar and grind all
Even, it is transferred in crucible；B) in Muffle furnace, crucible is heated to 700 DEG C, keeps this temperature 10h, after being subsequently cooled to room temperature
With nitric acid and the deionized water thoroughly cleaning of 1M, remove impurity, in atmosphere 80 DEG C of drying, collect strontium titanates nano-particle and treat
With.
Described step 2) in, as follows to the preprocess method of glass-carbon electrode：Glass-carbon electrode is used successively 0.3 μm of particle diameter and
After 0.5 μm of alumina powder polishing, then the alumina powder of deionized water flushing residual, put into ultrasonic clear in diluted nitric acid aqueous solution
Wash, finally clean glass-carbon electrode successively with ethanol and redistilled water, quickly dried up with nitrogen before modifying.
A kind of method that electrochemical glucose biology sensor using the present invention is detected to glucose sample, including
Following steps：
(1) in test fluid, with prepared electrochemica biological sensor as working electrode, saturated calomel electrode is as reference
Electrode, platinum plate electrode, as to electrode, adds glucose sample；
(2) electrochemical signals of working electrode and then with electrochemical workstation are detected.
In step (1), testing conditions are：Described test fluid is phosphate-buffered salt 0.02-0.1mol L-1Solution；Institute
State phosphate-buffered salt PBS solution, its pH value is 6.5-7.5.
Beneficial effects of the present invention：Sensor of the invention preparation is simple, quick, low cost, sensitivity height, stability
Good, can be used for the detection of concentration of glucose.And, using strontium titanates nano-particle fixing glucose oxidase, can be fine
Ground keeps the biology catalytic activity of large biological molecule.This sensor detects concentration of glucose by cyclic voltammetry, can be answered
For glucose electrochemical analysis.The invention has the advantages that：
1. the present invention adopts the strontium titanates nano-particle of preparation under different condition, and strontium titanates nano material has specific surface area
Greatly, the advantages of good biocompatibility, good conductivity, using its fixing glucose oxidase, be conducive to keeping the biologically active of enzyme,
Improve electron transport rate during detection, widen the range of linearity of detection.The nano material electrochemical glucose sensor of preparation
Can achieve the quantitative determination to glucose, sensitivity is high, good stability.
2nd, the preparation method of electrochemical glucose biology sensor of the present invention, preparation is simple, quick, low cost, sensitivity
Height, can be used for the detection of concentration of glucose in actual sample.This sensor is detected by the i t curve that ampere detection obtains
Concentration of glucose, can be applied to glucose electrochemical analysis.
3rd, it is placed in detection architecture using sensor of the invention, add glucose, detect its electrochemical signals.Test side
Method need not mark, simply, quickly, low cost, sensitivity is high, reappearance and good stability, can be used for blood sugar for human body glucose
The detection of concentration.In order to improve the accuracy of detection, the present invention also coating strontium titanates nano material mixed solution before, by glass
Carbon electrode uses the alumina powder polishing of 0.3 μm and 0.5 μm of particle diameter successively, then the alumina powder falling residual with deionized water rinsing
Afterwards, put in diluted nitric acid aqueous solution and be cleaned by ultrasonic, finally clean glass-carbon electrode successively with ethanol and redistilled water, use before modifying
Nitrogen quickly dries up, and the glass-carbon electrode of pretreatment is obtained with this.
4th, detection method, is optimized to the current potential in the pH of cushioning liquid, ampere detection, and obtains
Good glucose detection condition, improves analytical effect.The method is simple, quick, low cost, sensitivity are high, reappearance and steady
Qualitative good, for detecting human blood glucose concentration, contribute to the detection of diabetes, be that human health provides safeguard.
Brief description
Fig. 1 is that the solution (B) of strontium titanates nano-particle (A), strontium titanates nano-particle and shitosan modifies glass-carbon electrode
Upper dried scanning electron microscope (SEM) photograph (SEM)；
Fig. 2 is GOD/SrTiO3/ CS modified electrode, in 0.1M pH 7.0PBS, is continuously added to glucose variable concentrations
The i-t ampere response curve obtaining；
Specific embodiment
Below the experimentation of the present invention is described in detail it is intended to make the design cycle of the present invention, purpose of design
And its innovative point and advantage more clear.
First, preparation technology：
Embodiment 1
The present invention provides a kind of preparation method of strontium titanates nano material, by strontium titanates nano material and glucose oxidase
Modify on glass-carbon electrode, prepared electrochemica biological sensor.
The preparation method of the electrochemical glucose biology sensor of strontium titanates nano-particle of the present invention, step is such as
Under：
(1) strontium titanate nanoparticles are prepared, as follows：
A) strontium dioxide of the amount of certain material, the titanium dioxide (P25) of the amount of certain material and equimolar chlorine are weighed
Change sodium and potassium chloride, mix in mortar and grind uniformly, be transferred in crucible；
B) in Muffle furnace, crucible is heated to 700 DEG C, keeps this temperature 10h, after being subsequently cooled to room temperature, use the nitre of 1M
Acid and deionized water thoroughly cleaning, remove impurity, in atmosphere 80 DEG C of drying, collect strontium titanates nano-particle stand-by.
(2) under ultrasound condition, by 1.0-3.0mg strontium titanates nano-particle (SrTiO3) it is dispersed in 1.0mL's 0.5%
In shitosan (CS) solution, form strontium titanates nano-particle/shitosan mixed solution.
(3) glass-carbon electrode is used successively the alumina powder polishing of 0.3 μm and 0.5 μm of particle diameter, then fallen with deionized water rinsing
After the alumina powder of residual, put in diluted nitric acid aqueous solution and be cleaned by ultrasonic, finally clean glass with ethanol and redistilled water successively
Carbon electrode, is quickly dried up with nitrogen before modifying, and the glass-carbon electrode of pretreatment is obtained.
(4) add glucose oxidase so as to mix in strontium titanates nano-particle/shitosan mixed solution, join
Glucose oxidase processed/strontium titanates nano-particle/shitosan mixed solution is (i.e.：GOD/SrTiO3/ CS mixed solution).Then will
Prepared containing glucose oxidase/strontium titanates nano-particle/shitosan (GOD/SrTiO3/ CS) mixed solution 3.0-7.0 μ
L is homogeneously applied to glass-carbon electrode (d=2-4mm) surface pre-processing, and is placed under 4 DEG C of temperature environment, is obtained after being dried
GOD/SrTiO3/ CS modified electrode.
Side glucose sample detected using the electrochemical glucose biology sensor that aforementioned preparation process obtains
Method, comprises the following steps：
(1) in test fluid, with prepared modified electrode as working electrode, saturated calomel electrode is as reference electrode, platinum
Plate electrode, as to electrode, adds glucose sample；
(2) electrochemical signals of working electrode and then with electrochemical workstation are detected.
Described detection method, in step (1), testing conditions are：Described test fluid is phosphate-buffered salt 0.02-
0.1mol L-1Solution (i.e. PBS solution)；Described phosphate-buffered salt PBS solution, its pH value is 6.5-7.5.
The present invention controls the activity of enzyme using the property of nano material, using SrTiO3Nano-particle and shell gather
The bio-compatibility of sugar constructs a kind of biology sensor.Following description of tests, the present invention is based on GOD/SrTiO3/ CS modifies
The biology sensor of GCE not only has good catalysis activity to glucose, and repeatability and stability are fine.
Embodiment 2
Embodiment 2 is with the difference of embodiment 1：Prepare strontium titanate nanoparticles, as follows：
A) strontium dioxide of the amount of certain material, the titanium dioxide (P25) of the amount of certain material and equimolar chlorine are weighed
Change sodium and potassium chloride, mix in mortar and grind uniformly, be transferred in crucible (strontium dioxide, the amount of the material of titanium dioxide
Ratio be 2:1,1:1,1:2)；
B) in Muffle furnace, crucible is heated to 700 DEG C, keeps this temperature 10h, after being subsequently cooled to room temperature, use the nitre of 1M
Acid and deionized water thoroughly cleaning, remove impurity, in atmosphere 80 DEG C of drying, collect strontium titanates nano-particle stand-by.
(2) under ultrasound condition, by 1.0-3.0mg strontium titanates nano-particle (SrTiO3) it is dispersed in 1.0mL's 0.5%
In shitosan (CS) solution, form strontium titanates nano-particle/shitosan mixed solution.
(3) glass-carbon electrode is used successively the alumina powder polishing of 0.3 μm and 0.5 μm of particle diameter, then fallen with deionized water rinsing
After the alumina powder of residual, put in diluted nitric acid aqueous solution and be cleaned by ultrasonic, finally clean glass with ethanol and redistilled water successively
Carbon electrode, is quickly dried up with nitrogen before modifying, and the glass-carbon electrode of pretreatment is obtained.
(4) add glucose oxidase so as to mix in strontium titanates nano-particle/shitosan mixed solution, join
Glucose oxidase processed/strontium titanates nano-particle/shitosan mixed solution is (i.e.：GOD/SrTiO3/ CS mixed solution).Then will
Prepared containing glucose oxidase/strontium titanates nano-particle/shitosan (GOD/SrTiO3/ CS) mixed solution 3.0-7.0 μ
L is homogeneously applied to glass-carbon electrode (d=2-4mm) surface pre-processing, and is placed under 4 DEG C of temperature environment, is obtained after being dried
GOD/SrTiO3/ CS modified electrode.
Side glucose sample detected using the electrochemical glucose biology sensor that aforementioned preparation process obtains
Method, comprises the following steps：
(1) in test fluid, with prepared modified electrode as working electrode, saturated calomel electrode is as reference electrode, platinum
Plate electrode, as to electrode, adds glucose sample；
(2) electrochemical signals of working electrode and then with electrochemical workstation are detected.
Described detection method, in step (1), testing conditions are：Described test fluid is phosphate-buffered salt 0.02-
0.1mol L-1Solution (i.e. PBS solution)；Described phosphate-buffered salt PBS solution, its pH value is 6.5-7.5.
Embodiment 3
Embodiment 3 is with the difference of embodiment 1：Prepare strontium titanate nanoparticles, as follows：
A) strontium dioxide of the amount of certain material, the titanium dioxide (P25) of the amount of certain material and equimolar chlorine are weighed
Change sodium and potassium chloride, mix in mortar and grind uniformly, be transferred in crucible (strontium dioxide, the amount of the material of titanium dioxide
Ratio be 1：1)；
B) in Muffle furnace, crucible is heated to 800 DEG C, keeps this temperature 10h, after being subsequently cooled to room temperature, use the nitre of 1M
Acid and deionized water thoroughly cleaning, remove impurity, in atmosphere 80 DEG C of drying, collect strontium titanates nano-particle stand-by.
(2) under ultrasound condition, by 1.0-3.0mg strontium titanates nano-particle (SrTiO3) it is dispersed in 1.0mL's 0.5%
In shitosan (CS) solution, form strontium titanates nano-particle/shitosan mixed solution.
(3) glass-carbon electrode is used successively the alumina powder polishing of 0.3 μm and 0.5 μm of particle diameter, then fallen with deionized water rinsing
After the alumina powder of residual, put in diluted nitric acid aqueous solution and be cleaned by ultrasonic, finally clean glass with ethanol and redistilled water successively
Carbon electrode, is quickly dried up with nitrogen before modifying, and the glass-carbon electrode of pretreatment is obtained.
(4) add glucose oxidase so as to mix in strontium titanates nano-particle/shitosan mixed solution, join
Glucose oxidase processed/strontium titanates nano-particle/shitosan mixed solution is (i.e.：GOD/SrTiO3/ CS mixed solution).Then will
Prepared containing glucose oxidase/strontium titanates nano-particle/shitosan (GOD/SrTiO3/ CS) mixed solution 3.0-7.0 μ
L is homogeneously applied to glass-carbon electrode (d=2-4mm) surface pre-processing, and is placed under 4 DEG C of temperature environment, is obtained after being dried
GOD/SrTiO3/ CS modified electrode.
Side glucose sample detected using the electrochemical glucose biology sensor that aforementioned preparation process obtains
Method, comprises the following steps：
(1) in test fluid, with prepared modified electrode as working electrode, saturated calomel electrode is as reference electrode, platinum
Plate electrode, as to electrode, adds glucose sample；
(2) electrochemical signals of working electrode and then with electrochemical workstation are detected.
Described detection method, in step (1), testing conditions are：Described test fluid is phosphate-buffered salt 0.02-
0.1mol L-1Solution (i.e. PBS solution)；Described phosphate-buffered salt PBS solution, its pH value is 6.5-7.5.
2nd, the Product characteristics made：
Referring to Fig. 1, Fig. 1 (A) is the SEM figure of the strontium titanates using present invention preparation, and Fig. 1 (B) is then to be disperseed with shitosan
The SEM figure of strontium titanates, as seen from the figure, shitosan with dispersed strontium titanates, and can form firm film.Shitosan has
The features such as preferable film forming ability and biocompatibility, nontoxicity, good mechanical strength and water penetration, it has been widely used in
Dispersion nano material and fixing protein molecules, thus build biology sensor.
Fig. 2 is GOD/SrTiO3/ CS/GCE is continuously stirring, connect in the PBS of the 0.1M pH 7.0 of the saturation of the air
The i-t ampere response curve that the glucose solution of continuous dropping variable concentrations obtains.Voltage is -0.5V, interior illustration：Response signal with
The relation of concentration of glucose.The concentration of peak current (Ipa, μ A) and glucose sugar is 1 × 10-5-1.2×10-3mol·L-1In the range of
In good linear relationship, detection is limited to 3 μM (S/N=3).The equation of its relation curve is y=0.04755+0.0011x, spirit
Sensitivity is 15.6mA M-1·cm-2, Michaelis constant is 0.1mM.
Claims (6)
1. a kind of electrochemical glucose biology sensor based on strontium titanates nano-particle is it is characterised in that be by grape glycosyloxy
Change enzyme and this electrochemical glucose biology sensor is obtained with strontium titanates Nanoparticle Modified in glass-carbon electrode.
2. a kind of preparation method of the electrochemical glucose biology sensor based on strontium titanates nano-particle it is characterised in that：Will
Glucose oxidase and strontium titanates Nanoparticle Modified, in glass-carbon electrode, are obtained this electrochemical glucose biology sensor.
3. method according to claim 2 is it is characterised in that inclusion step is as follows：
1) strontium titanates nano-particle is distributed in chitosan solution, adds glucose oxidase, be configured to grape glycoxidative
Enzyme/strontium titanates nano-particle/shitosan mixed solution；
2) glucose oxidase/strontium titanates nano-particle/shitosan mixed solution is uniformly coated to the glass carbon through pretreatment
Electrode surface, is placed in after being dried under 4 DEG C of temperature environment, that is, obtains described electrochemical glucose biology sensor.
4. according to claim 3 preparation method it is characterised in that：Described step 1) in glucose oxidase/strontium titanates
Nano-particle/shitosan mixed solution obtains by the following method：Under ultrasound condition, by 1.0-3.0mg strontium titanates nanoparticle
Son is scattered in 0.5% chitosan solution of 1.0mL, then the glucose oxidase of 10-20mg is added strontium titanates nanoparticle
Son/chitosan solution, is slowly stirred 10 minutes under room temperature.
5. preparation method according to claim 3 it is characterised in that：Described step 2) in strontium titanates nano-particle/grape
The consumption of carbohydrate oxidase/shitosan mixed solution is 3.0-7.0 μ L.
6. preparation method according to claim 3 it is characterised in that：Step 2) used in pre-process glass-carbon electrode place
Reason method is as follows：Glass-carbon electrode is used successively the alumina powder polishing of 0.3 μm and 0.5 μm of particle diameter, then deionized water flushing is residual
After the alumina powder staying, put in diluted nitric acid aqueous solution and be cleaned by ultrasonic, finally clean glass carbon with ethanol and redistilled water successively
Electrode, is quickly dried up with nitrogen before modifying.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104826158A (en) * | 2015-05-06 | 2015-08-12 | 南京文钧医疗科技有限公司 | Drug carrier with integrated functions and preparation method thereof |
| CN105401260A (en) * | 2015-11-03 | 2016-03-16 | 浙江大学 | Preparation method of strontium titanate nanotube material |
| CN105548317A (en) * | 2016-01-07 | 2016-05-04 | 河南科技学院 | Preparation method of electrochemical glucose biosensor and detection method for testing glucose |
-
2016
- 2016-09-29 CN CN201610864894.8A patent/CN106442674A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104826158A (en) * | 2015-05-06 | 2015-08-12 | 南京文钧医疗科技有限公司 | Drug carrier with integrated functions and preparation method thereof |
| CN105401260A (en) * | 2015-11-03 | 2016-03-16 | 浙江大学 | Preparation method of strontium titanate nanotube material |
| CN105548317A (en) * | 2016-01-07 | 2016-05-04 | 河南科技学院 | Preparation method of electrochemical glucose biosensor and detection method for testing glucose |
Non-Patent Citations (3)
| Title |
|---|
| HUI-LING LI 等: "Low temperature molten salt synthesis of SrTiO3 submicron crystallites and nanocrystals in the eutectic NaCl–KCl", 《MATERIALS LETTERS》 * |
| SEONG KEUN KIM 等: "Electric-field-induced Shift in the Threshold Voltage in LaAlO3/SrTiO3 Heterostructures", 《SCIENTIFIC REPORTS》 * |
| XIAOHUA CHEN 等: "Nanoplated bismuth titanate sub-microspheres for protein immobilization and their corresponding direct electrochemistry and electrocatalysis", 《BIOSENSORS AND BIOELECTRONICS》 * |
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Application publication date: 20170222 |