CN100483122C - Full blood lactic biological sensor modified by oriented carbon nano tube - Google Patents
Full blood lactic biological sensor modified by oriented carbon nano tube Download PDFInfo
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- CN100483122C CN100483122C CNB2005100492859A CN200510049285A CN100483122C CN 100483122 C CN100483122 C CN 100483122C CN B2005100492859 A CNB2005100492859 A CN B2005100492859A CN 200510049285 A CN200510049285 A CN 200510049285A CN 100483122 C CN100483122 C CN 100483122C
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- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 title claims abstract description 32
- 239000002041 carbon nanotube Substances 0.000 title claims abstract description 32
- 229910021393 carbon nanotube Inorganic materials 0.000 title claims abstract description 32
- 210000004369 blood Anatomy 0.000 title claims abstract description 23
- 239000008280 blood Substances 0.000 title claims abstract description 23
- 239000002245 particle Substances 0.000 claims abstract description 10
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 9
- 239000003381 stabilizer Substances 0.000 claims abstract description 9
- 238000009413 insulation Methods 0.000 claims description 11
- 238000013016 damping Methods 0.000 claims description 9
- 239000012530 fluid Substances 0.000 claims description 9
- 108050006365 L-lactate oxidases Proteins 0.000 claims description 8
- -1 potassium ferricyanide Chemical compound 0.000 claims description 8
- 238000000151 deposition Methods 0.000 claims description 7
- 230000000694 effects Effects 0.000 claims description 7
- 238000000034 method Methods 0.000 claims description 6
- 238000002848 electrochemical method Methods 0.000 claims description 5
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 claims description 4
- RBTBFTRPCNLSDE-UHFFFAOYSA-N 3,7-bis(dimethylamino)phenothiazin-5-ium Chemical compound C1=CC(N(C)C)=CC2=[S+]C3=CC(N(C)C)=CC=C3N=C21 RBTBFTRPCNLSDE-UHFFFAOYSA-N 0.000 claims description 4
- 108091003079 Bovine Serum Albumin Proteins 0.000 claims description 4
- 229920002101 Chitin Polymers 0.000 claims description 4
- 108010022355 Fibroins Proteins 0.000 claims description 4
- 241000233855 Orchidaceae Species 0.000 claims description 4
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 claims description 4
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 claims description 4
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 claims description 4
- 229910052782 aluminium Inorganic materials 0.000 claims description 4
- 239000004411 aluminium Substances 0.000 claims description 4
- 229940098773 bovine serum albumin Drugs 0.000 claims description 4
- 238000005229 chemical vapour deposition Methods 0.000 claims description 4
- 229960000907 methylthioninium chloride Drugs 0.000 claims description 4
- 239000008363 phosphate buffer Substances 0.000 claims description 4
- 239000011148 porous material Substances 0.000 claims description 4
- 238000004544 sputter deposition Methods 0.000 claims description 4
- 230000003197 catalytic effect Effects 0.000 claims description 3
- 230000008021 deposition Effects 0.000 claims description 3
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 abstract description 18
- 235000014655 lactic acid Nutrition 0.000 abstract description 9
- 239000004310 lactic acid Substances 0.000 abstract description 9
- 230000035945 sensitivity Effects 0.000 abstract description 3
- 230000008901 benefit Effects 0.000 abstract description 2
- 238000005516 engineering process Methods 0.000 abstract description 2
- 239000007788 liquid Substances 0.000 abstract description 2
- 239000000203 mixture Substances 0.000 abstract description 2
- 102000004316 Oxidoreductases Human genes 0.000 abstract 1
- 108090000854 Oxidoreductases Proteins 0.000 abstract 1
- 230000003139 buffering effect Effects 0.000 abstract 1
- 239000003054 catalyst Substances 0.000 abstract 1
- 238000002203 pretreatment Methods 0.000 abstract 1
- 239000000758 substrate Substances 0.000 abstract 1
- JVTAAEKCZFNVCJ-REOHCLBHSA-N L-lactic acid Chemical compound C[C@H](O)C(O)=O JVTAAEKCZFNVCJ-REOHCLBHSA-N 0.000 description 8
- 230000004044 response Effects 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 238000002484 cyclic voltammetry Methods 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
- 239000004020 conductor Substances 0.000 description 2
- 239000003792 electrolyte Substances 0.000 description 2
- 239000008151 electrolyte solution Substances 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- ICFJFFQQTFMIBG-UHFFFAOYSA-N phenformin Chemical compound NC(=N)NC(=N)NCCC1=CC=CC=C1 ICFJFFQQTFMIBG-UHFFFAOYSA-N 0.000 description 2
- 229960003243 phenformin Drugs 0.000 description 2
- 239000000276 potassium ferrocyanide Substances 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- XOGGUFAVLNCTRS-UHFFFAOYSA-N tetrapotassium;iron(2+);hexacyanide Chemical compound [K+].[K+].[K+].[K+].[Fe+2].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-] XOGGUFAVLNCTRS-UHFFFAOYSA-N 0.000 description 2
- 208000010444 Acidosis Diseases 0.000 description 1
- 229920002134 Carboxymethyl cellulose Polymers 0.000 description 1
- 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 description 1
- JVTAAEKCZFNVCJ-UHFFFAOYSA-M Lactate Chemical compound CC(O)C([O-])=O JVTAAEKCZFNVCJ-UHFFFAOYSA-M 0.000 description 1
- 108010073450 Lactate 2-monooxygenase Proteins 0.000 description 1
- 230000007950 acidosis Effects 0.000 description 1
- 208000026545 acidosis disease Diseases 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 239000001768 carboxy methyl cellulose Substances 0.000 description 1
- 235000010948 carboxy methyl cellulose Nutrition 0.000 description 1
- 239000008112 carboxymethyl-cellulose Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 230000034659 glycolysis Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical compound C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 description 1
- 239000002071 nanotube Substances 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
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- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The full blood lactic acid biosensor modified with oriented carbon nanotube features that one layer of Co particles as the catalyst for oriented growth of carbon nanotube is deposited on the substrate and that the reaction layer has base layer comprising oriented carbon nanotube as the second electric intermediate and the upper layer comprising L-lactic oxidase and the mixture of reagent, stabilizer and buffering liquid as the first electric intermediate. Compared with available the technology, the present invention has the advantages of obviously raised responding sensitivity on lactic acid concentration of the sensor, work voltage of the sensor lowered from 500 mv to 300 mv, no need of pre-treatment of the blood sample and blood sample amount as less as 2 microliter.
Description
Technical field
The present invention relates to a kind of biology sensor, particularly a kind of full blood lactic biological sensor of modified by oriented carbon nano tube.
Background technology
The full blood lactic biological sensor of modified by oriented carbon nano tube 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 this sensor and measure the order of severity that lactic acid content in the blood of human body is used to disclose some disease, as 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 full blood lactic biological sensor of modified by oriented carbon nano tube, comprise base plate 1, electrode 2, cavity 3, responding layer 4, cover plate 5, it is characterized in that: electrode 2 is the Au electrodes that form on base plate 1 with vacuum sputtering, cavity 3 usefulness plastic sheetings surround around covering electrode 2, on an one sidewall, have the pore 3a that a sample introduction is used, responding layer 4 is arranged in the cavity 3, and the bottom 4a of responding layer is as the aligned carbon nanotube layer of second electron medium, and the aligned carbon nanotube layer is to utilize electrochemical method at Al
20
3Deposition Co particle is as catalyzer in the vertical micropore of insulation course, and adopt chemical vapour deposition technique to obtain, the oriented growth time is 5 minutes, be 4～8 hours ultrasonic break time, the upper strata 4b of responding layer is the potpourri that drips the L-Lactate Oxidase and form as reagent, stabilizing agent, the damping fluid of first electron medium, and dry; Cover plate 5 is the plastic sheetings that cover on the cavity 3, have an air hole on it, contain 500～1000 unit of activity of L-Lactate Oxidase in the responding layer 4, reagent is selected a kind of in the potassium ferricyanide, methylene blue, the Prussia orchid for use, and its content is 40～200ug, and stabilizing agent is selected a kind of in trehalose, chitin, fibroin albumen, the bovine serum albumin(BSA) for use, its content is 20～80ug, damping fluid is a phosphate buffer, and content is 60～240ug, and its pH value is 6.0～7.0.Above-mentioned base plate 1 is that its surface forms Al through anodized
2O
3The aluminium sheet of insulation course.
With prior art relatively, the present invention has following outstanding advantage: 1) 1 aligned carbon nanotube in responding layer makes that as second electron medium potassium ferrocyanide (the reduction form of the first mediator potassium ferricyanide) is easy to give directed carbon nano-tube with electron transport, aligned carbon nanotube can be given electrode with electron transport again apace under the low voltage effect, thereby significantly improves the response sensitivity of L-lactic acid concn; 2) the while aligned carbon nanotube can effectively improve the activity of enzyme, impels reaction to carry out fast; 3) nanotubes in the aligned carbon nanotube can effectively be stored water of crystallization, 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 drawing of the full blood lactic biological sensor of modified by oriented carbon nano tube.
Fig. 2 is the parts vertical view of the full blood lactic biological sensor of modified by oriented carbon nano tube.
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 nano-tube, the reversibility of potassium ferricyanide reaction improves, and potassium ferrocyanide is easy to give carbon nano-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 aligned carbon nanotube, the response sensitivity of sensor is improved and improves.
Embodiment
Embodiment 1:
A kind of full blood lactic biological sensor of modified by oriented carbon nano tube, base plate 1 are that its surface forms Al through anodized
2O
3The aluminium sheet of insulation course, and adopt electrochemical method at Al
2O
3Deposition is as the grown Co particle of catalytic action of aligned carbon nanotube in vertical micropore of insulation course; Electrode 2 is the Au electrodes that form on base plate 1 with vacuum sputtering, cavity 3 usefulness plastic sheetings surround around covering electrode 2, on an one sidewall, have the pore 3a that a sample introduction is used, responding layer 4 is arranged in the cavity 3, its bottom 4a is the aligned carbon nanotube layer as second electron medium, the aligned carbon nanotube layer adopts chemical vapour deposition technique to obtain, the oriented growth time is 5 minutes, be 4 hours ultrasonic break time, its upper strata 4b drips the L-Lactate Oxidase and as the reagent of first electron medium, stabilizing agent, the potpourri that damping fluid is formed, and dry; Cover plate 5 is the plastic sheetings that cover on the cavity 3, has an air hole on it; Contain 500 unit of activity of L-Lactate Oxidase in the responding layer 4, reagent is selected a kind of in the potassium ferricyanide, methylene blue, the Prussia orchid for use, its content is 40ug, stabilizing agent is selected a kind of in trehalose, chitin, fibroin albumen, the bovine serum albumin(BSA) for use, its content is 20ug, damping fluid is a phosphate buffer, and its pH value is 6.0, and content is 60ug.The Co particle of deposition is to be the CoSO of 60g/l with concentration
47H
2O solution and concentration are the H of 3.5g/l
3BO
3Solution is made into electrolytic solution, controls its proportioning, and making electrolyte ph is 3.1, adopts electrochemical process that the Co particle is deposited on Al
2O
3On the insulation course.
Embodiment 2:
A kind of full blood lactic biological sensor of modified by oriented carbon nano tube, base plate 1 are that its surface forms Al through anodized
2O
3The aluminium sheet of insulation course, and adopt electrochemical method at Al
2O
3Deposition is as the grown Co particle of catalytic action of aligned carbon nanotube in vertical micropore of insulation course; Electrode 2 is the Au electrodes that form on base plate 1 with vacuum sputtering, cavity 3 usefulness plastic sheetings surround around covering electrode 2, on an one sidewall, have the pore 3a that a sample introduction is used, responding layer 4 is arranged in the cavity 3, its bottom 4a is the aligned carbon nanotube layer as second electron medium, the aligned carbon nanotube layer adopts chemical vapour deposition technique to obtain, the oriented growth time is 5 minutes, be 8 hours ultrasonic break time, the reagent that its upper strata 4b drips the L-Lactate Oxidase and uses as first electron medium, stabilizing agent, the potpourri that damping fluid is formed, and dry; Cover plate 5 is the plastic sheetings that cover on the cavity 3, has an air hole on it; L-contains 1000 unit of activity of Lactate Oxidase in the responding layer 4, reagent is selected a kind of in the potassium ferricyanide, methylene blue, the Prussia orchid for use, its content is 200ug, stabilizing agent is selected a kind of in trehalose, chitin, fibroin albumen, the bovine serum albumin(BSA) for use, its content is 80ug, damping fluid is a phosphate buffer, and its pH value is 7.0, and content is 240ug.The Co particle of deposition is to be the CoSO of 60g/l with concentration
47H
2O solution and concentration are the H of 3.5g/l
3BO
3Solution is made into electrolytic solution, controls its proportioning, and making electrolyte ph is 3.7, adopts electrochemical process that the Co particle is deposited on Al
2O
3On the insulation course.
List of references
[1]Nobuo?Shimojo，Keiichi?Naka，Harumi?Uenoyama，et，al，Electrochemical?Assay?Systemwith?Sing-leUse?Electrode?Strip?for?Measuring?Lactate?in?Whole?Blood，ClinicalChemistry，Vol，39，No，11，1993
Claims (1)
1. the full blood lactic biological sensor of a modified by oriented carbon nano tube comprises base plate (1), electrode (2), and cavity (3), responding layer (4), cover plate (5) is characterized in that: base plate (1) is that its surface forms Al through anodized
2O
3The aluminium sheet of insulation course, and adopt electrochemical method at Al
2O
3Deposition is as the grown Co particle of catalytic action of aligned carbon nanotube in vertical micropore of insulation course; Electrode (2) is to go up the Au electrode that forms at base plate (1) with vacuum sputtering, cavity (3) covers electrode (2) with plastic sheeting and surrounds on every side, on an one sidewall, have the pore (3a) that a sample introduction is used, responding layer (4) is arranged in the cavity (3), the bottom of responding layer (4a) is the aligned carbon nanotube layer of second electron medium, and the aligned carbon nanotube layer is to utilize electrochemical method at Al
2O
3The Co particle of deposition is as catalyzer in the vertical micropore of insulation course, and adopt chemical vapour deposition technique to obtain, the oriented growth time is 5 minutes, be 4～8 hours ultrasonic break time, the upper strata of responding layer (4b) is the potpourri that drips the L-Lactate Oxidase and form as reagent, stabilizing agent, the damping fluid of first electron medium, and dry; Cover plate (5) is the plastic sheeting that covers on the cavity (3), have an air hole on it, contain 500～1000 unit of activity of L-Lactate Oxidase in the responding layer (4), reagent is selected a kind of in the potassium ferricyanide, methylene blue, the Prussia orchid for use, and its content is 40～200ug, and stabilizing agent is selected a kind of in trehalose, chitin, fibroin albumen, the bovine serum albumin(BSA) for use, its content is 20～80ug, damping fluid is a phosphate buffer, and content is 60～240ug, and its pH value is 6.0～7.0.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100492859A CN100483122C (en) | 2005-01-31 | 2005-01-31 | Full blood lactic biological sensor modified by oriented carbon nano tube |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100492859A CN100483122C (en) | 2005-01-31 | 2005-01-31 | Full blood lactic biological sensor modified by oriented carbon nano tube |
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| Publication Number | Publication Date |
|---|---|
| CN1648650A CN1648650A (en) | 2005-08-03 |
| CN100483122C true CN100483122C (en) | 2009-04-29 |
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Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1987454B (en) * | 2006-10-31 | 2011-01-12 | 成都夸常医学工业有限公司 | Nano structure component containing oriented nano structure functional zone and its preparing and applying method |
| LU101273B1 (en) * | 2019-06-21 | 2020-12-28 | Ecole Polytechnique Fed Lausanne Epfl | Sensing platform |
| CN112240900B (en) * | 2020-10-21 | 2022-05-06 | 南京工业大学 | Preparation method of lactic acid biosensor |
-
2005
- 2005-01-31 CN CNB2005100492859A patent/CN100483122C/en not_active Expired - Fee Related
Non-Patent Citations (2)
| Title |
|---|
| 氧化铝模板上定向纳米碳管的快速生长及超声切短. 徐军明，张孝彬，陈飞，李昱，王幼文.物理化学学报，第20卷第3期. 2004 |
| 氧化铝模板上定向纳米碳管的快速生长及超声切短. 徐军明，张孝彬，陈飞，李昱，王幼文.物理化学学报，第20卷第3期. 2004 * |
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