CN102243208A - Preparation method for glucose biosensor based on ternary hydrotalcite and derivative thereof - Google Patents
Preparation method for glucose biosensor based on ternary hydrotalcite and derivative thereof Download PDFInfo
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- CN102243208A CN102243208A CN2011101566713A CN201110156671A CN102243208A CN 102243208 A CN102243208 A CN 102243208A CN 2011101566713 A CN2011101566713 A CN 2011101566713A CN 201110156671 A CN201110156671 A CN 201110156671A CN 102243208 A CN102243208 A CN 102243208A
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- glucose
- ldh
- mgnife
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Abstract
The invention relates to a preparation method for glucose biosensors based on ternary hydrotalcite (LDH) and derivatives thereof. According to the invention, excellent characteristics of nano-materials of LDH and its derivatives are utilized to effectively fix glucose oxidase (GOD) and promote electron transfer in the method. The method comprises the following steps: uniformly dispersing 1.6mg nano-materials of MgNiFe-LDH and Mg(Ni)Fe2O4 in 1 mL of a chitosam (CHT) solution with a concentration of 0.5% (w/v) by ultrasonic dispersion, adding 1.6 mg of GOD, and continuing ultrasonic dispersion of the mixture so as to obtain desired modification agents; dropwisely applying the modification agents on the surface of glassy carbon electrodes so as to obtain the glucose biosensors. According to the invention, nano-materials of MgNiFe-LDH and Mg(Ni)Fe2O4 are respectively used to fix GOD, and composite membranes of MgNiFe-LDH/GOD/CHT and Mg(Ni)Fe2O4/GOD/CHT are respectively used to modify the glassy carbon electrodes so as to prepare two glucose biosensors for electrochemical gauging of glucose; enzyme fixed on the glucose biosensors has high activity, good stability and strong specificity, and determination of glucose with the glucose biosensors has the characteristics of rapidness, sensitivity, accuracy, etc.
Description
Technical field
The present invention relates to utilize two kinds of nano material hydrotalcite MgNiFe-LDH and derivative Mg(Ni thereof) Fe
2O
4Fixing glucose oxidase and with complex film M gNiFe-LDH/GOD/CHT and Mg(Ni) Fe
2O
4/ GOD/CHT modified glassy carbon electrode is made the method for glucose biological sensor, belongs to nanometer technology and electrochemical analysis detection technique field.
Background technology
In recent years, along with the raising of people's living standard and increasing of elderly population, diabetes become international frequently-occurring disease and common disease, and serious danger coercing human health, becomes the third-largest dangerous disease that is only second to cardiovascular disease and cancer.Therefore the mensuration of the content of glucose has great significance to prevention and the treatment of diabetes.
Electrochemica biological sensor since its simple, quick, highly sensitive and good selective in glucose detection, occupy an important position.Glucose oxidase becomes the ideal model that people prepare glucose sensor because of its catalytic efficiency height, good stability and to the characteristics such as catalysis height selectivity of D-glucose.Since first glucose sensor of preparation in 1967, glucose biological sensor has experienced the first generation, the second generation and third generation biology sensor from Updike and Hicks.And the electron mediator that artificial adding has the catalytic oxidation reducing action is big characteristics of second generation biology sensor.The present invention promptly utilizes ferrocenecarboxylic acid as electron mediator.
But (easily run off, it is permanent etc. that activity can not keep) limited the application of this type of glucose sensor because some weakness of enzyme.Therefore effectively fixing glucose oxidase becomes the research emphasis of this type of glucose sensor to keep enzymatic activity.The researcher utilizes multiple material such as polymer film for this reason, and mesoporous material, CNT and clay etc. fix the activity that enzyme keeps enzyme.
Anionic clay claims that again (molecular formula is [M (II) to hydrotalcite (LDHs)
1-
y M (III)
y (OH)
2]
y+
[(X
y/
n n-
)
mH
2O]
y-
) because of its special poriness, water percentage height, and nontoxic, the low and structural stability of cost waits physics and chemical property well, for immobilized enzyme provides coenocorrelation preferably, especially by Co
2+, Ni
2+, Mn
3+Deng the hydrotalcite that the transition metal that has electron transport and redoxomorphism in aqueous slkali is formed, caused the interest of more and more researchers recently.As NiAl-LDH, hydrotalcites such as ZnAl-LDH have been used to prepare biology sensor.But the hydrotalcite MgNiFe-LDH that three elements forms and derivative thereof also are not used in this type of research.The present invention is exactly the research that utilizes synthetic nano material MgNiFe-LDH of coprecipitation and derivant fixing glucose oxidase thereof to be used for glucose biological sensor.
The present invention combines nanometer technology and biosensor technique.For biology enzyme provides good coenocorrelation, make glucose oxidase in this environment, give full play to catalytic action by nano material hydrotalcite or derivatives thereof to glucose.
Summary of the invention
Purpose of the present invention is providing two kinds of new nano material hydrotalcite MgNiFe-LDH and derivative Mg(Ni thereof) Fe
2O
4, and utilize the effective fixing glucose oxidase of good characteristic (GOD) of nano material, and the synergy of the shitosan good with film forming (CHT) has been prepared dressing agent MgNiFe-LDH/GOD/CHT and Mg(Ni) Fe
2O
4/ GOD/CHT composite membrane drips the method that glass-carbon electrode has been made a kind of glucose biological sensor that is coated with.
A kind of preparation method of the glucose biological sensor based on ternary hydrotalcite and derivant thereof is characterized in that having following preparation process and step:
A. at first prepare ternary hydrotalcite crystal MgNiFe-LDH and derivative Mg(Ni thereof) Fe
2O
4, with the coprecipitation preparation, its preparation process is as follows: prepare earlier ternary hydrotalcite crystal MgNiFe-LDH, adding mol ratio under inert atmosphere in three-necked bottle is the Mg (NO of 2:1:1
3)
26H
2O, Ni (NO
3)
26H
2O and Fe (NO
3)
39H
2The O aqueous solution is heated to 40 ℃ under vigorous stirring, drip a certain proportion of NaOH and Na
2CO
3Alkali lye mixed liquor (2:1), until pH value of solution=10, gained suspension continues to stir 100 ℃ of lower crystallization after 6 hours then, suction filtration, washing are to neutral, the gained filter cake is dry 24 h in 70 ℃ baking oven, namely get the ternary hydrotalcite nano particle, are designated as MgNiFe-LDH;
MgNiFe-LDH 900 ℃ of lower roastings 2 hours, is namely prepared hydrotalcite derivant, i.e. nano material Mg(Ni) Fe
2O
4
B. with ternary hydrotalcite MgNiFe-LDH and derivative Mg(Ni thereof) Fe
2O
4Nano material respectively and the combination of shitosan (CHT), glucose oxidase (GOD) be modified at and be prepared into two kinds of biology sensors that are used for measuring concentration of glucose on the glass-carbon electrode; Its preparation process is as follows:
B-1. process glass-carbon electrode with traditional method: at first with the abrasive paper for metallograph polishing, use successively then 1.0 μ m, 0.3 μ m and 0.05 μ m A1
2O
3Suspension is polished to minute surface at chamois leather, uses respectively at last HNO
3Solution (1:1), absolute ethyl alcohol and intermediate water ultrasonic cleaning;
B-2. the preparation of dressing agent: with ternary hydrotalcite MgNiFe-LDH or the derivative Mg(Ni of 1.6 mg) Fe
2O
4Nano material be scattered in 1 mL CHT acetum (0.5%,
w/
v, i.e. g/mL), ultrasonic being uniformly dispersed adds 1.6 mg GOD and continues ultrasonic to being uniformly dispersed;
B-3. above-mentioned two kinds of finely dispersed dressing agent 10 μ L are dripped and be coated on the clean glass-carbon electrode surface, dried overnight in 4 ℃ of refrigerators makes electrode surface form uniform MgNiFe-LDH/GOD/CHT or Mg(Ni) Fe
2O
4/ GOD/CHT composite membrane; Namely get based on nano material ternary hydrotalcite MgNiFe-LDH or derivatives thereof Mg(Ni) Fe
2O
4Glucose biological sensor.
A kind of mensuration of concentration of glucose adopts the above-mentioned glucose biological sensor based on ternary hydrotalcite and derivant thereof to measure, and its feature has following determination step and is:
A. before using, this chemical modification glass-carbon electrode is soaked in the phosphate buffered solution of pH=7.0 earlier, then-0.4～+ potential window of 0.8V in cyclic voltammetry to be scanned up to figure stable; Rinse well with redistilled water again;
B. with nano material complex film MgNiFe-LDH/GOD/CHT or Mg(Ni) Fe
2O
4The glass-carbon electrode that/GOD/CHT modifies is as working electrode, and saturated calomel electrode is reference electrode, and platinum plate electrode is to electrode, forms traditional three-electrode system; During electrochemical gaging, modified electrode is placed in the phosphate buffer of the 0.1 M pH 7.0 that stirs with constant rate of speed, has added 0.5 mM ferrocenecarboxylic acid in the buffer solution; Under 0.60 V and the best current potential of 0.50 V, record current-time graph when background current reaches stable state, adds the glucose sample with microsyringe, the record current response signal respectively; Obtain the dense response time-current curve of glucose; Record concentration of glucose according to Conventional Calculation Method then.。
The present invention utilizes respectively ternary hydrotalcite MgNiFe-LDH or derivatives thereof ferrospinel Mg(Ni) Fe
2O
4The peculiar property of nano particle comes effective fixing glucose oxidase, and with complex film M gNiFe-LDH/GOD/CHT or Mg(Ni) Fe
2O
4/ GOD/CHT drips and is coated with glass-carbon electrode, prepares the glucose oxidase enzyme biologic sensor.Its advantage is: the specific surface area of (1) nano particle is big, has meso-hole structure, and the efficient height of immobilized enzyme between nanoparticle surface or hole has effectively kept the activity of enzyme; (2) layer structure of hydrotalcite has promoted the electron transport effect between enzyme and electrolyte mediator; Hydrotalcite derivant is that spinel type ferrite has cube structure, and it is a soft magnetic ferrite, has certain dispersibility the reaction between enzyme and the matrix is accelerated, and detection limit is lower.
Modified electrode among the present invention is the electrochemical sensor of a class novel nano-material immobilized enzyme, the fast electrochemical that is used for glucose is measured, utilize that hydrotalcite and the prepared glucose biological sensor of derivates nanometer particle thereof have active height, good stability, cost is low and the life-span such as grows at advantage, be expected to obtain practical application.
Description of drawings
Fig. 1 a, b are respectively the hydrotalcite MgNiFe-LDH(a that has layer structure among the present invention) and derivative-spinel type ferrite Mg(Ni) Fe
2O
4(b) x-ray diffraction pattern of nano material (abscissa is X-ray scanning angle 2T, and ordinate is X-ray diffraction peak intensity I).
Fig. 2 a, b are respectively among the present invention the respectively complex film M gNiFe-LDH/GOD/CHT(a that forms with shitosan of fixing glucose oxidase of two kinds of nano materials) and Mg(Ni) Fe
2O
4/ GOD/CHT(b) the glucose biological sensor that makes up of modified glassy carbon electrode, (abscissa is time t, and ordinate is current value to the time current curve of concentration of glucose response under optimal condition
I p).
Embodiment
After now specific embodiments of the invention being described in.
The preparation process of biology sensor and step are as follows in this enforcement:
A. at first prepare ternary hydrotalcite crystal MgNiFe-LDH and derivative Mg(Ni thereof) Fe
2O
4, with the coprecipitation preparation, its preparation process is as follows: prepare earlier ternary hydrotalcite crystal MgNiFe-LDH, adding mol ratio under inert atmosphere in three-necked bottle is the Mg (NO of 2:1:1
3)
26H
2O, Ni (NO
3)
26H
2O and Fe (NO
3)
39H
2The O aqueous solution is heated to 40 ℃ under vigorous stirring, drip a certain proportion of NaOH and Na
2CO
3Alkali lye mixed liquor, volume ratio are 2:1, until pH value of solution=10, gained suspension continue to stir down 100 ℃ of lower crystallization after 6 hours then, and suction filtration, washing be to neutral, gained filter cake dry 24 h in 70 ℃ baking oven, namely get the ternary hydrotalcite nano particle, be designated as MgNiFe-LDH.MgNiFe-LDH 900 ℃ of lower roastings 2 hours, is namely prepared hydrotalcite derivant, i.e. nano material Mg(Ni) Fe
2O
4
B. biology sensor preparation:
(1) at first polishes glass-carbon electrode with abrasive paper for metallograph, use successively then 1.0 μ m, 0.3 μ m and 0.05 μ m A1
2O
3Suspension is polished to minute surface at deerskin, uses respectively at last HNO
3(1:1), absolute ethyl alcohol and intermediate water ultrasonic cleaning;
(2) preparation of dressing agent and sensor: with 1.6 mg ternary hydrotalcite MgNiFe-LDH or derivative Mg(Ni) Fe
2O
4Nano material is scattered in acetic acid (0.5%, w/v the is g/mL) solution of 1 mL CHT, and ultrasonic being uniformly dispersed adds 1.6 mg GOD and continue ultrasonic to being uniformly dispersed; With the finely dispersed dressing agent MgNiFe-LDH/GOD/CHT of 10 μ L or Mg(Ni) Fe
2O
4/ GOD/CHT drips and is coated on the clean glass-carbon electrode surface, and dried overnight in 4 ℃ of refrigerators makes electrode surface form uniform MgNiFe-LDH/GOD/CHT or Mg(Ni) Fe
2O
4/ GOD/CHT composite membrane namely gets based on nano material ternary hydrotalcite MgNiFe-LDH or derivatives thereof Mg(Ni) Fe
2O
4Glucose biological sensor.
Utilize the method for above-mentioned biosensor assay concentration of glucose, its step is as follows:
When (1) using, earlier chemically modified electrode is soaked in the phosphate buffered solution of pH=7.0, cyclic voltammetry is scanned up to stable in the potential window of-0.4～0.8V then; Rinse well with redistilled water again.
(2) glass-carbon electrode of above-mentioned chemical modification is used for the mensuration of concentration of glucose; Concrete assay method is as follows: as working electrode, saturated calomel electrode is a contrast electrode with the nano material complex film modified glassy carbon, and the platinized platinum electrode is to electrode, forms traditional three-electrode system; During electrochemical gaging, modified electrode is placed in the phosphate buffer of the 0.1 M pH 7.0 that stirs with constant rate of speed, has added 0.5 mM ferrocenecarboxylic acid in the damping fluid.Be under 0.60 V and 0.5 V at current potential respectively, record current-time curve when background current reaches stable state, adds the glucose sample with microsyringe, the record current response signal; Obtain concentration of glucose response time-current curve, record concentration of glucose according to conventional computing method then.
The chemical property of nanometer material structure and chemically modified electrode characterizes:
By x-ray diffractometer figure, synthesized as can be known hydrotalcite material MgNiFe-LDH with good crystalline structure and the spinel type ferrite material Mg(Ni of cubic crystal structure) Fe
2O
4, such as Fig. 1 a, shown in the b.
Chemically modified electrode carries out electrochemical gaging to glucose
Under best test condition, MgNiFe-LDH/GOD/CHT(a) or Mg(Ni) Fe
2O
4/ GOD/CHT(b) complex film modified electrode pair glucose electric current-time response as shown in Figure 2.As seen from the figure, reach the time all very short (all less than 10 s) of 95% steady-state current, this mainly is because the nano material that makes has good characteristics such as biocompatibility, poriness and electron transport promotion property to biology enzyme.Prepared biology sensor all increases along with the increase of concentration of glucose gradually to the current-responsive of glucose.The complex film modified electrode of MgNiFe-LDH/GOD/CHT is in 1.0-8.0 mM scopes, and electric current and concentration of glucose are linear, and linearly dependent coefficient r=0.989 is detected and is limited to 0.12 mM (signal to noise ratio is 3); Use Mg(Ni) Fe
2O
4The complex film modified electrode of/GOD/CHT is in 1.0-7.0mM scope, and electric current and concentration of glucose are linear, and linearly dependent coefficient r=0.991 is detected and is limited to 0.05 mM (signal to noise ratio is 3).
Based on nano material ternary hydrotalcite MgNiFe-LDH or derivatives thereof Mg(Ni) Fe
2O
4Glucose biological sensor showed good reappearance and stability.Isocyatic glucose solution is carried out 15 times measure, its standard deviation is respectively 1.165% and 2.18%.Electrode is not after the time spent deposits one month in 4 ℃ refrigerator, and current-responsive all remains on more than 90%.And be not subjected to the interference of some common metallic ions or other carbohydrate such as fructose, wood sugar, selectivity is good.
Claims (2)
1. preparation method based on the glucose biological sensor of ternary hydrotalcite and derivant thereof is characterized in that having following preparation process and step:
A. at first prepare ternary hydrotalcite crystal MgNiFe-LDH and derivative Mg(Ni thereof) Fe
2O
4, with the coprecipitation preparation, its preparation process is as follows: prepare earlier ternary hydrotalcite crystal MgNiFe-LDH, adding mol ratio under inert atmosphere in three-necked bottle is the Mg (NO of 2:1:1
3)
26H
2O, Ni (NO
3)
26H
2O and Fe (NO
3)
39H
2The O aqueous solution is heated to 40 ℃ under vigorous stirring, drip a certain proportion of NaOH and Na
2CO
3Alkali lye mixed liquor (2:1), until pH value of solution=10, gained suspension continues to stir 100 ℃ of lower crystallization after 6 hours then, suction filtration, washing are to neutral, the gained filter cake is dry 24 h in 70 ℃ baking oven, namely get the ternary hydrotalcite nano particle, are designated as MgNiFe-LDH;
MgNiFe-LDH 900 ℃ of lower roastings 2 hours, is namely prepared hydrotalcite derivant, i.e. nano material Mg(Ni) Fe
2O
4
B. with ternary hydrotalcite MgNiFe-LDH and derivative Mg(Ni thereof) Fe
2O
4Nano material respectively and the combination of shitosan (CHT), glucose oxidase (GOD) be modified at and be prepared into two kinds of biology sensors that are used for measuring concentration of glucose on the glass-carbon electrode; Its preparation process is as follows:
B-1. process glass-carbon electrode with traditional method: at first with the abrasive paper for metallograph polishing, use successively then 1.0 μ m, 0.3 μ m and 0.05 μ m A1
2O
3Suspension is polished to minute surface at chamois leather, uses respectively at last HNO
3Solution (1:1), absolute ethyl alcohol and intermediate water ultrasonic cleaning;
B-2. the preparation of dressing agent: with ternary hydrotalcite MgNiFe-LDH or the derivative Mg(Ni of 1.6 mg) Fe
2O
4Nano material be scattered in 1 mL CHT acetum (0.5%,
w/
v, i.e. g/mL), ultrasonic being uniformly dispersed adds 1.6 mg GOD and continues ultrasonic to being uniformly dispersed;
B-3. above-mentioned two kinds of finely dispersed dressing agent 10 μ L are dripped and be coated on the clean glass-carbon electrode surface, dried overnight in 4 ℃ of refrigerators makes electrode surface form uniform MgNiFe-LDH/GOD/CHT or Mg(Ni) Fe
2O
4/ GOD/CHT composite membrane; Namely get based on nano material ternary hydrotalcite MgNiFe-LDH or derivatives thereof Mg(Ni) Fe
2O
4Glucose biological sensor.
2. the mensuration of a concentration of glucose adopts and measures according to the described glucose biological sensor based on ternary hydrotalcite and derivant thereof of claim, and its feature has following determination step and is:
A. before using, this chemical modification glass-carbon electrode is soaked in the phosphate buffered solution of pH=7.0 earlier, then-0.4～+ potential window of 0.8V in cyclic voltammetry to be scanned up to figure stable; Rinse well with redistilled water again;
B. with nano material complex film MgNiFe-LDH/GOD/CHT or Mg(Ni) Fe
2O
4The glass-carbon electrode that/GOD/CHT modifies is as working electrode, and saturated calomel electrode is reference electrode, and platinum plate electrode is to electrode, forms traditional three-electrode system; During electrochemical gaging, modified electrode is placed in the phosphate buffer of the 0.1 M pH 7.0 that stirs with constant rate of speed, has added 0.5 mM ferrocenecarboxylic acid in the buffer solution; Under 0.60 V and the best current potential of 0.50 V, record current-time graph when background current reaches stable state, adds the glucose sample with microsyringe, the record current response signal respectively; Obtain the dense response time-current curve of glucose; Record concentration of glucose according to Conventional Calculation Method then.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109432450A (en) * | 2018-11-23 | 2019-03-08 | 北京化工大学 | A kind of supermolecule nano-chemistry dynamical medicine and its application in terms of oncotherapy |
| CN111289593A (en) * | 2020-02-28 | 2020-06-16 | 北京农业信息技术研究中心 | Microelectrode array sensor for in-vivo detection of plant glucose and preparation and application thereof |
| CN111289592A (en) * | 2020-02-28 | 2020-06-16 | 北京农业信息技术研究中心 | Microarray sensor for simultaneously detecting various saccharides in plant body in vivo and preparation and application thereof |
| CN112542328A (en) * | 2020-12-02 | 2021-03-23 | 浙江大学 | Ternary layered metal hydroxide @ polyaniline composite electrode material and preparation method and application thereof |
| CN113092553A (en) * | 2021-03-03 | 2021-07-09 | 哈尔滨工业大学（深圳） | Novel method for detecting glucose by enzyme-free glucose sensor |
| CN114216945A (en) * | 2021-12-14 | 2022-03-22 | 安徽大学 | Nickel-iron oxide composite material and preparation method and application thereof |
-
2011
- 2011-06-13 CN CN2011101566713A patent/CN102243208A/en active Pending
Non-Patent Citations (2)
| Title |
|---|
| YANHONG XU 等: "《A Glucose Sensor Based on a Nano-Structured Metal Composite Oxide Derived from Hydrotalcite》", 《SENSOR LETTERS》 * |
| YANHONG XU 等: "《Preparation and electrochemical investigation of a nano-structured material Ni2+/MgFe layered double hydroxide as a glucose biosensor》", 《APPLIED CLAY SCIENCE》 * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109432450A (en) * | 2018-11-23 | 2019-03-08 | 北京化工大学 | A kind of supermolecule nano-chemistry dynamical medicine and its application in terms of oncotherapy |
| CN111289593A (en) * | 2020-02-28 | 2020-06-16 | 北京农业信息技术研究中心 | Microelectrode array sensor for in-vivo detection of plant glucose and preparation and application thereof |
| CN111289592A (en) * | 2020-02-28 | 2020-06-16 | 北京农业信息技术研究中心 | Microarray sensor for simultaneously detecting various saccharides in plant body in vivo and preparation and application thereof |
| CN111289593B (en) * | 2020-02-28 | 2022-07-19 | 北京农业信息技术研究中心 | Microelectrode array sensor for in-vivo detection of plant glucose and preparation and application thereof |
| CN111289592B (en) * | 2020-02-28 | 2022-07-22 | 北京农业信息技术研究中心 | Microarray sensor for simultaneously detecting various saccharides in plant body in vivo and preparation and application thereof |
| CN112542328A (en) * | 2020-12-02 | 2021-03-23 | 浙江大学 | Ternary layered metal hydroxide @ polyaniline composite electrode material and preparation method and application thereof |
| CN113092553A (en) * | 2021-03-03 | 2021-07-09 | 哈尔滨工业大学（深圳） | Novel method for detecting glucose by enzyme-free glucose sensor |
| CN114216945A (en) * | 2021-12-14 | 2022-03-22 | 安徽大学 | Nickel-iron oxide composite material and preparation method and application thereof |
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Application publication date: 20111116 |