CN1187608C - biosensor for lactic acid in whole blood - Google Patents
biosensor for lactic acid in whole blood Download PDFInfo
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- CN1187608C CN1187608C CNB031294332A CN03129433A CN1187608C CN 1187608 C CN1187608 C CN 1187608C CN B031294332 A CNB031294332 A CN B031294332A CN 03129433 A CN03129433 A CN 03129433A CN 1187608 C CN1187608 C CN 1187608C
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Abstract
The present invention relates to a full blood lactic acid biological sensor. The full blood glucose biological sensor is characterized in that a cavity body (3) is formed by covering a plastic film around an electrode (2), and a capillary bore communicated with the cavity body is arranged on the cavity body; a reaction layer (4) comprises carboxymethyl cellulose CMC (4a), a decorative carbon nanometer tube (4b) used as a second electronic intermediate, L-lactic acid oxidase and mixtures (4c) which is composed of a reagent, a stabilizing agent and buffer liquid and is used as a first electronic intermediate, wherein the carboxymethyl cellulose, the carbon nanometer tube, the L-lactic acid oxidase and the mixed liquid are orderly overlapped in the cavity body. Compared with the prior art, the full blood glucose biological sensor has the advantages that because the decorative carbon nanometer tube as the second electronic intermediate is added to the reaction layer, the response sensitivity to L-lactic acid concentration is obviously enhanced, the voltage applied to the electrode is 300 mv when the L-lactic acid content in blood is measured, the applied voltage of the prior art is 0.5v, a sample inlet pore of the cavity body is positioned at a lateral wing of the sensor, fingers are used for clamping both sides of the electrode when the sample is introduced, the sensor deformation can not occur, the volume change of the blood sucked into the cavity body is avoided, few blood samples are needed by the sensor, 2 mu l of blood is only needed, and the blood samples do not need to be pretreated when measured.
Description
Technical field
The present invention relates to a kind of biology sensor, particularly a kind of full blood lactic biological sensor.
Background technology
Full blood lactic biological sensor is a kind of biology sensor that utilizes biomolecule to measure fast, accurately, easily lactic acid content in the liquid such as blood as sensitive element, use the order of severity that lactic acid content in this sensor measurement blood of human body is used to disclose some disease, be used to instruct the diabetic suitably to take phenformin, the excessive use phenformin can cause glycolysis, lactic acid content is raise, cause acidosis, jeopardize patient's life; Be used to measure sportsman's concentration of lactic acid, scientifically controlled motion person's training strength reaches the optimum training effect; In Food Science, can determine the freshness of food by detecting concentration of lactic acid.At present large-scale biochemical analyzer is mainly adopted in the measurement of lactic acid, price is very expensive.Described lactic acid content is meant the L-lactic acid content.
In amperometric biosensor, there is the people to adopt single electron medium to add oxidasic mode and is developed into disposable L-lactic biological sensor [1], this sensor is made up of PVC base plate, electrode, cavity, responding layer, cavity cover plate, wherein electrode and contact conductor adopt homogenous material to make, responding layer is the responding layer of carboxymethyl cellulose, L-Lactate Oxidase, the potassium ferricyanide (electronic conduction body), damping fluid composition, and the sample holes of cavity is opened at the electrode other end.The defective of this sensor is: the voltage that is added on the contact conductor is higher, is 0.5v, and antijamming capability is not good enough, and its reason is that the potassium ferricyanide is directly relatively poor at the redox characteristic of carbon electrodes, needs to improve voltage and obtains response current preferably.Chinese patent CN1219676A has introduced the two enzyme biologic sensors of a kind of disposable bielectron mediator, be used for detecting cholesterol in the blood, glucose etc., but this sensor needs the participation of oxygen, is subjected to the interference that oxygen content changes in the blood easily.
Summary of the invention
A kind of whole blood L-lactic biological sensor, comprise PVC base plate 1, electrode 2, cavity 3, responding layer 4, cavity cover plate 5, responding layer 4 is arranged in the cavity 3, it is characterized in that: electrode 2 is carbon electrodes of making on base plate 1 with screen printing technique, and contact conductor 2a is the silver wire of making on base plate 1 with screen printing technique; Cavity 3 covered with plastic film around electrode 2 forms, a horizontal sample holes that communicates with cavity is arranged on it, this sample holes is a pore, responding layer 4 comprises carboxymethyl cellulose 4a, modification carbon nanometer tube 4b as electronics second mediator, this modification carbon nanometer tube be modified with-COOH ,-carbon nanometer tube of OH, its content is 4～20ug, the potpourri 4c that L-Lactate Oxidase, the reagent as first electron medium, stabilizing agent, damping fluid are formed, layering is superimposed upon on the carbon electrode successively, and successively dry respectively; Have an air hole on the cavity cover plate 5, carboxymethyl cellulose 4a content is 10～40ug, contains 1～2 unit of activity of L-Lactate Oxidase in the responding layer 4; The first electron medium reagent in the responding layer 4 is selected a kind of in the potassium ferricyanide, methylene blue, the Prussia orchid for use, and content is 40～200ug; Stabilizing agent in the responding layer 4 is selected a kind of in trehalose, chitin, fibroin albumen, the bovine serum albumin(BSA) for use, and content is 20～80ug; Damping fluid in the responding layer 4 is a phosphate buffer, and its pH value is 6.0～7.0, and content is 60～240ug.
With prior art relatively, the present invention has following outstanding advantage: 1) the modification carbon nanometer tube in responding layer makes that as second electron medium potassium ferrocyanide (the reduction form of the first mediator potassium ferricyanide) is easy to give carbon nanometer tube with electron transport, carbon nanometer tube can be given electrode with electron transport apace under the low voltage effect, thereby significantly improves the response sensitivity of L-lactic acid concn; 2) the while carbon nanometer tube can effectively improve the activity of enzyme, impels reaction to carry out fast; 3) water of crystallization can effectively be stored in the sodium mitron road in the carbon nanometer tube, helps the long preservation and the stability of enzyme; When 4) using in this sensor measurement blood the L-lactic acid content, the voltage that is added on the electrode is 300mv, and the voltage that prior art applies is 0.5v; 5) sample holes of cavity is positioned at the sensor flank, and fingerhold electrode both sides can not cause the sensor distortion during sample introduction, avoid sucking the change of cavity inner blood amount; 6) blood sample of this sensor needs is few, does not need blood sample is carried out pre-service during only need 2 μ l, and measurement.
Description of drawings
Fig. 1 is the structural representation of full blood lactic biological sensor.
Fig. 2 is the vertical view of base plate 1, electrode 2, contact conductor 2a, cavity 3, cavity cover plate 5.
Fig. 3 is the cyclic voltammetry curve figure of electrode, and A is the cyclic voltammetry curve of this sensor among the figure, and B is the cyclic voltammetry curve of existing sensor.After showing the adding electronics second mediator carbon nanometer tube, the reversibility of potassium ferricyanide reaction improves, and potassium ferrocyanide is easy to give carbon nanometer tube with electron transport.
Fig. 4 is the current-responsive graph of a relation of this sensor to L-lactic acid content in the blood, and C is this sensor current response relation among the figure, and D is the current-responsive relation of existing sensor.After showing the adding carbon nanometer tube, the response sensitivity of sensor is improved and improves.
Embodiment
Embodiment 1:
A kind of full blood lactic biological sensor, as shown in Figure 1, use screen printing technique and on PVC base plate 1, make thick film carbon electrode 2 and silver contact conductor 2a respectively, covered with plastic film forms cavity 3 around carbon electrode 2, splashing into 2 μ l concentration in cavity is 5mg/mlCMC solution, solution forms CMC layer 4a behind 60 ℃ of oven dryings; Point adds 2 μ l and modifies carbon nanometer tube suspending liquid, its concentration is 2mg/ml, behind 60 ℃ of oven dryings, form carbon nanometer tube layer 4b, select again and add 2 μ l mixed liquors, composition is that 500 unit of activity/ml of L-Lactate Oxidase, potassium ferricyanide 50mg/ml, seaweeds 20mg/ml, pH value are 6.0, concentration is the phosphate buffer of 0.2M, form mixolimnion 4c after the drying case drying, on cavity 2, cover the volume that layer of plastic film is determined cavity 3 subsequently, this volume is 2 μ l, and plastic sheeting herein is cavity cover plate 5, has air hole on it.
Embodiment 2:
A kind of full blood lactic biological sensor, as shown in Figure 1, use screen printing technique and on PVC base plate 1, make thick film carbon electrode 2 and silver contact conductor 2a respectively, covered with plastic film forms cavity 3 around carbon electrode 2, splashing into 2 μ l concentration in cavity is the cmc soln of 20mg/ml, solution forms CMC layer 4a behind 60 ℃ of oven dryings; Drip 2 μ l and modify carbon nanometer tube suspending liquid, its concentration is 6mg/ml, behind 60 ℃ of oven dryings, form carbon nanometer tube layer 4b, select again and add 2 μ l mixed liquors, composition is that 1000 unit of activity/ml of L-Lactate Oxidase, potassium ferricyanide 50mg/ml, bovine serum albumin(BSA) 4mg/ml, pH value are 7.0, concentration is the phosphate buffer of 0.4M, form mixolimnion 4c after the drying case drying, on cavity 3, cover layer of plastic film subsequently, as cavity cover plate 5, the volume of cavity 3 is 2 μ l, has air hole on the cavity cover plate 5.
List of references
[1]Nobuo?Shimojo，Keiichi?Naka，Harumi?Uenoyama，et，al，Electrochemical?Assay?System?withSing-leUse?Electrode?Strip?for?Measuring?Lactate?in?Whole?Blood，Glinical?Chemistry，Vol，39，No，11，1993
Claims (6)
1. full blood lactic biological sensor, comprise PVC base plate (1), electrode (2), cavity (3), responding layer (4), cavity cover plate (5), responding layer (4) is arranged in the cavity (3), it is characterized in that: electrode (2) is the carbon electrode of making on base plate (1) with screen printing technique, and contact conductor (2a) is the silver wire of making on base plate (1) with serigraphy; Cavity (3) is in electrode (2) covered with plastic film formation on every side, a horizontal sample holes that communicates with cavity is arranged on it, this sample holes is a pore, responding layer (4) comprises carboxymethyl cellulose (4a), modification carbon nanometer tube (4b) as second electron medium, this modification carbon nanometer tube be modified with-COOH ,-carbon nanometer tube of OH, its content is 4～20ug, the mixed liquor (4c) that L-Lactate Oxidase, the reagent as electronics first mediator, stabilizing agent, damping fluid are formed layering successively is superimposed upon on the carbon electrode, and successively dry respectively; Have an air hole on the cavity cover plate (5).
2. biology sensor according to claim 1 is characterized in that: its content of carboxymethyl cellulose (4a) is 10～40ug.
3. biology sensor according to claim 1 is characterized in that: contain 1～2 unit of activity of L-Lactate Oxidase in the responding layer (4).
4. biology sensor according to claim 1 is characterized in that: the reagent in the responding layer (4) is selected a kind of in the potassium ferricyanide, methylene blue, the Prussia orchid for use, and its content is 40～200ug.
5. biology sensor according to claim 1 is characterized in that: the stabilizing agent in the responding layer (4) is selected a kind of in trehalose, chitin, fibroin albumen, the bovine serum albumin(BSA) for use, and its content is 20～80ug.
6. biology sensor according to claim 1 is characterized in that: the damping fluid in the responding layer (4) is a phosphate buffer, and its pH value is 6.0～7.0, and its content is 60～240ug.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB031294332A CN1187608C (en) | 2003-06-18 | 2003-06-18 | biosensor for lactic acid in whole blood |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB031294332A CN1187608C (en) | 2003-06-18 | 2003-06-18 | biosensor for lactic acid in whole blood |
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| Publication Number | Publication Date |
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| CN1462880A CN1462880A (en) | 2003-12-24 |
| CN1187608C true CN1187608C (en) | 2005-02-02 |
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| CNB031294332A Expired - Fee Related CN1187608C (en) | 2003-06-18 | 2003-06-18 | biosensor for lactic acid in whole blood |
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Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| TWI336782B (en) | 2007-07-05 | 2011-02-01 | Apex Biotechnology Corp | Composite modified electrode trip |
| CN101349667B (en) * | 2007-07-16 | 2011-11-23 | 中国科学院化学研究所 | Electrochemical detection device of physiological active substance and special-purpose electrochemical sensor preparation method thereof |
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