CN111239228A - Electrochemical biosensor and method for measuring blood impedance phase angle - Google Patents
Electrochemical biosensor and method for measuring blood impedance phase angle Download PDFInfo
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- CN111239228A CN111239228A CN202010110724.7A CN202010110724A CN111239228A CN 111239228 A CN111239228 A CN 111239228A CN 202010110724 A CN202010110724 A CN 202010110724A CN 111239228 A CN111239228 A CN 111239228A
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- 239000008280 blood Substances 0.000 title claims abstract description 26
- 210000004369 blood Anatomy 0.000 title claims abstract description 26
- 238000000034 method Methods 0.000 title claims abstract description 17
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 16
- 239000000758 substrate Substances 0.000 claims abstract description 14
- 239000007788 liquid Substances 0.000 claims abstract description 4
- 239000004094 surface-active agent Substances 0.000 claims description 10
- 239000007853 buffer solution Substances 0.000 claims description 7
- 239000010408 film Substances 0.000 claims description 6
- LZZYPRNAOMGNLH-UHFFFAOYSA-M Cetrimonium bromide Chemical compound [Br-].CCCCCCCCCCCCCCCC[N+](C)(C)C LZZYPRNAOMGNLH-UHFFFAOYSA-M 0.000 claims description 5
- 102000004190 Enzymes Human genes 0.000 claims description 5
- 108090000790 Enzymes Proteins 0.000 claims description 5
- MOTZDAYCYVMXPC-UHFFFAOYSA-N dodecyl hydrogen sulfate Chemical compound CCCCCCCCCCCCOS(O)(=O)=O MOTZDAYCYVMXPC-UHFFFAOYSA-N 0.000 claims description 5
- KTWOOEGAPBSYNW-UHFFFAOYSA-N ferrocene Chemical compound [Fe+2].C=1C=C[CH-]C=1.C=1C=C[CH-]C=1 KTWOOEGAPBSYNW-UHFFFAOYSA-N 0.000 claims description 5
- -1 potassium ferricyanide Chemical compound 0.000 claims description 5
- 239000001397 quillaja saponaria molina bark Substances 0.000 claims description 5
- 229930182490 saponin Natural products 0.000 claims description 5
- 150000007949 saponins Chemical class 0.000 claims description 5
- 239000003381 stabilizer Substances 0.000 claims description 5
- GPRLSGONYQIRFK-MNYXATJNSA-N triton Chemical compound [3H+] GPRLSGONYQIRFK-MNYXATJNSA-N 0.000 claims description 5
- GBDZMMXUOBAJMN-UHFFFAOYSA-K azane;ruthenium(3+);trichloride Chemical compound N.N.N.N.N.N.[Cl-].[Cl-].[Cl-].[Ru+3] GBDZMMXUOBAJMN-UHFFFAOYSA-K 0.000 claims description 4
- FHCPAXDKURNIOZ-UHFFFAOYSA-N tetrathiafulvalene Chemical compound S1C=CSC1=C1SC=CS1 FHCPAXDKURNIOZ-UHFFFAOYSA-N 0.000 claims description 4
- 239000010409 thin film Substances 0.000 claims description 4
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 claims description 3
- 229910052709 silver Inorganic materials 0.000 claims description 3
- 239000004332 silver Substances 0.000 claims description 3
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims description 2
- 229910052737 gold Inorganic materials 0.000 claims description 2
- 239000010931 gold Substances 0.000 claims description 2
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical compound C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 claims description 2
- 239000012491 analyte Substances 0.000 description 7
- 238000001514 detection method Methods 0.000 description 5
- 238000005534 hematocrit Methods 0.000 description 4
- 238000000840 electrochemical analysis Methods 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 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
- DDRJAANPRJIHGJ-UHFFFAOYSA-N creatinine Chemical compound CN1CC(=O)NC1=N DDRJAANPRJIHGJ-UHFFFAOYSA-N 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 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
- KJTLSVCANCCWHF-UHFFFAOYSA-N Ruthenium Chemical compound [Ru] KJTLSVCANCCWHF-UHFFFAOYSA-N 0.000 description 1
- LEHOTFFKMJEONL-UHFFFAOYSA-N Uric Acid Chemical compound N1C(=O)NC(=O)C2=C1NC(=O)N2 LEHOTFFKMJEONL-UHFFFAOYSA-N 0.000 description 1
- TVWHNULVHGKJHS-UHFFFAOYSA-N Uric acid Natural products N1C(=O)NC(=O)C2NC(=O)NC21 TVWHNULVHGKJHS-UHFFFAOYSA-N 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- 229940109239 creatinine Drugs 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 229940043264 dodecyl sulfate Drugs 0.000 description 1
- 238000000835 electrochemical detection Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000003891 environmental analysis Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- XEOSBIMHSUFHQH-UHFFFAOYSA-N fulvalene Chemical compound C1=CC=CC1=C1C=CC=C1 XEOSBIMHSUFHQH-UHFFFAOYSA-N 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 150000002576 ketones Chemical class 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000004451 qualitative analysis Methods 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 238000011897 real-time detection Methods 0.000 description 1
- 229910052707 ruthenium Inorganic materials 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 229940116269 uric acid Drugs 0.000 description 1
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/416—Systems
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/02—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/416—Systems
- G01N27/4163—Systems checking the operation of, or calibrating, the measuring apparatus
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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Electrochemistry (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
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Abstract
The invention discloses an electrochemical biosensor, which comprises a substrate layer, wherein the upper end of the substrate layer is provided with an electrode output end, the lower end of the substrate layer is provided with a reaction electrode and an auxiliary electrode, conductive circuits are connected among the reaction electrode, the auxiliary electrode and the electrode output end, a reaction channel at the lower end of the substrate layer is provided with a hydrophilic film layer, and the front end of the hydrophilic film layer is provided with a liquid inlet to be detected; the hydrophilic film layer at the rear end of the reaction channel is respectively provided with air holes, and the reaction electrode and the auxiliary electrode are provided with reagent layers. The invention also discloses a method for measuring the blood impedance phase angle by using the electrochemical biosensor.
Description
Technical Field
The invention belongs to the technical field of electrochemical detection and analysis and biosensors, and relates to an electrochemical biosensor for measuring an impedance phase angle of a whole blood sample and a method for measuring the impedance phase angle of blood.
Background
Biosensors are widely used in the fields of medical detection, environmental analysis, food detection, etc., and electrochemical biosensors are a technology for performing qualitative or quantitative analysis on an analyte by chemically reacting with the analyte in a certain manner and converting reaction signals into electrical signals that can be analyzed and measured. The sensor has the characteristics of simple operation, small sample amount, high accuracy, low manufacturing cost, capability of being used for real-time detection and the like, but the conventional electrochemical biosensor is easily influenced by the hematocrit of blood when testing the concentration of an analyte in a blood sample, so that the concentration result of the analyte is interfered. Most of the existing hematocrit test methods use a blood flow rate method, a conductance method or an impedance method to correct the analyte concentration. The methods are greatly influenced by the stability of a printing electrode of the sensor and the properties of a hydrophilic layer film material, so that the development of a new detection method for improving the sensitivity and the accuracy of a detection means is a main problem to be solved urgently.
Disclosure of Invention
In order to eliminate the influence of the hematocrit in the blood on the measurement of the concentration of the specific component in the blood and improve the detection accuracy. The invention provides an electrochemical biosensor and a method for measuring blood impedance phase angle, which can correct the measured analyte concentration of a whole blood sample through the blood sample parameters measured by the biosensor.
The technical scheme of the invention is as follows: an electrochemical biosensor comprises a substrate layer, wherein an electrode output end is arranged at the upper end of the substrate layer, a reaction electrode and an auxiliary electrode are arranged at the lower end of the substrate layer, conductive circuits are connected among the reaction electrode, the auxiliary electrode and the electrode output end, a hydrophilic thin film layer is arranged on a reaction channel at the lower end of the substrate layer, and a liquid inlet to be detected is arranged at the lower end of the hydrophilic thin film layer; the upper end of the hydrophilic film layer is provided with air holes, and the reaction electrode and the auxiliary electrode are provided with reagent layers.
Further, the buffer solution in the reagent layer maintains the pH value in the range of 6 to 8.
Further, the reagent layer comprises a surfactant, an electron mediator, an enzyme and a stabilizer.
Preferably, the electron mediator includes one of potassium ferricyanide, ruthenium hexaammonium trichloride, fulvalene tetrasulfide, and ferrocene.
Preferably, the surfactant comprises one or more of saponin, dodecyl sulfate, cetyl trimethyl ammonium bromide, and triton.
Further, the reaction electrode and the auxiliary electrode adopt: a combination of one or more of gold electrodes, carbon electrodes, silver electrodes.
The invention also discloses a method for measuring the blood impedance phase angle by utilizing the electrochemical biosensor, wherein the reagent layer comprises a surfactant, an electronic mediator, an enzyme and a stabilizer; the reaction is carried out under the condition that the constant potential between the reaction electrode and the auxiliary electrode is between 0.2 and 1.0V; the amplitude of the alternating current impedance of the alternating current voltage signals applied to the reaction electrode and the auxiliary electrode is 0.1-0.4V, and the frequency is 100-; the buffer solution in the reagent layer is used to maintain the pH in the range of 6 to 8.
Further to the method, the buffer solution in the reagent layer maintains the pH value in the range of 6 to 8.
Preferably, the electron mediator includes one of potassium ferricyanide, ruthenium, tetrathiafulvalene and ferrocene.
Preferably, the surfactant comprises one or more of saponin, dodecyl sulfuric acid, cetyl trimethyl ammonium bromide and triton.
Compared with the prior art, the invention has the following technical scheme characteristics:
(1) the invention provides a brand-new electrochemical testing method which is small in blood consumption, short in testing time, few in interference factors and convenient to operate.
(2) The electrochemical test strip has simple structure, is easy for large-scale production and has good industrial application prospect.
Drawings
FIG. 1 is a schematic structural diagram of an electrochemical biosensor for measuring blood impedance phase angle according to an embodiment of the present invention.
Detailed Description
In order that those skilled in the relevant art can understand the technical solution of the present invention, the technical solution in the embodiments of the present invention is completely and clearly described in conjunction with the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only some embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Fig. 1 is a schematic structural diagram of an electrochemical biosensor for measuring blood impedance phase angle according to the present embodiment, which includes a substrate layer 12, a conductive layer on the substrate layer 12, and an insulating layer 11 disposed above the conductive layer. One end of the conducting layer comprises 5 electrode output ends 1-5, the other end of the conducting layer is sequentially provided with a first auxiliary electrode 9, a working electrode 8, a reaction electrode 7 and a second auxiliary electrode 10, conductive circuits 6 are connected among the reaction electrode 7, the first auxiliary electrode 9, the working electrode 8, the second auxiliary electrode 10 and the electrode output ends 1-5, a hydrophilic film layer 14 is arranged on a reaction channel at the lower end of the base layer, and the lower end of the hydrophilic film layer 14 is provided with a liquid inlet 15 to be detected; the upper end of the hydrophilic film layer 14 is provided with an air hole 16, and the reaction electrode 7 and the second auxiliary electrode 10 are provided with a reagent layer 13.
The buffer solution in the reagent layer 13 maintains the pH value in the range of 6 to 8.
The reagent layer 13 includes a surfactant, an electron mediator, an enzyme, and a stabilizer. The electron mediator comprises one of potassium ferricyanide, hexaammonium ruthenium trichloride, tetrathiafulvalene and ferrocene. The surfactant comprises one or more of saponin, dodecyl sulfuric acid, cetyl trimethyl ammonium bromide and triton. The reaction electrode and the auxiliary electrode are silver electrodes.
The method for measuring the blood impedance phase angle by the electrochemical biosensor of the embodiment comprises the following measurement conditions: the reaction is carried out under the condition of constant potential between 0.2V and 1.0V between the reaction electrode 7 and the second auxiliary electrode 10; the amplitude of the AC impedance of the AC voltage signal applied to the reaction electrode 8 and the second auxiliary electrode 9 is 0.1-0.4V, and the frequency is 100-.
The electrochemical analysis method by measuring the impedance phase angle of the whole blood sample comprises the following steps: measuring the impedance phase angle of the blood sample, establishing a correlation curve of the impedance phase angle and the hematocrit value, and finally correcting the blood analyte concentration by using the correlation curve. The analytes are mainly glucose, total cholesterol, creatinine, blood ketone, uric acid and the like in whole blood.
The above embodiments are merely illustrative of the technical solutions and features of the present invention, and are intended to enable those skilled in the art to understand the contents of the present invention and implement the present invention, and not to limit the protection scope of the present invention. Various changes or modifications of the invention based on the principles thereof are also intended to be defined by the following claims.
Claims (10)
1. An electrochemical biosensor, comprising: the device comprises a substrate layer, wherein an electrode output end is arranged at the upper end of the substrate layer, a reaction electrode and an auxiliary electrode are arranged at the lower end of the substrate layer, conductive circuits are connected among the reaction electrode, the auxiliary electrode and the electrode output end, a hydrophilic thin film layer is arranged on a reaction channel at the lower end of the substrate layer, and a liquid inlet to be detected is arranged at the lower end of the hydrophilic thin film layer; the upper end of the hydrophilic film layer is provided with air holes, and the reaction electrode and the auxiliary electrode are provided with reagent layers.
2. The electrochemical biosensor according to claim 1, wherein:
the buffer solution in the reagent layer maintains the pH in the range of 6 to 8.
3. The electrochemical biosensor according to claim 1, wherein:
the reagent layer comprises a surfactant, an electron mediator, an enzyme and a stabilizer.
4. The electrochemical biosensor according to claim 3, wherein:
the electron mediator comprises one of potassium ferricyanide, hexaammonium ruthenium trichloride, tetrathiafulvalene and ferrocene.
5. The electrochemical biosensor according to claim 3, wherein:
the surfactant comprises one or more of saponin, dodecyl sulfuric acid, cetyl trimethyl ammonium bromide and triton.
6. The electrochemical biosensor according to claim 1, wherein:
the reaction electrode and the auxiliary electrode adopt: a combination of one or more of gold electrodes, carbon electrodes, silver electrodes.
7. A method of measuring a blood impedance phase angle using the electrochemical biosensor of claim 1, wherein: the reagent layer comprises a surfactant, an electron mediator, an enzyme and a stabilizer; the reaction is carried out under the condition that the constant potential between the reaction electrode and the auxiliary electrode is between 0.2 and 1.0V; the amplitude of the alternating current impedance of the alternating current voltage signals applied to the reaction electrode and the auxiliary electrode is 0.1-0.4V, and the frequency is 100-; the buffer solution in the reagent layer is used to maintain the pH in the range of 6 to 8.
8. The method of measuring blood impedance phase angle of claim 7 wherein:
the buffer solution in the reagent layer maintains the pH in the range of 6 to 8.
9. The method of measuring blood impedance phase angle of claim 7 wherein:
the electron mediator comprises one of potassium ferricyanide, hexaammonium ruthenium trichloride, tetrathiafulvalene and ferrocene.
10. The method of measuring blood impedance phase angle of claim 7 wherein:
the surfactant comprises one or more of saponin, dodecyl sulfuric acid, cetyl trimethyl ammonium bromide and triton.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010110724.7A CN111239228A (en) | 2020-02-24 | 2020-02-24 | Electrochemical biosensor and method for measuring blood impedance phase angle |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202010110724.7A CN111239228A (en) | 2020-02-24 | 2020-02-24 | Electrochemical biosensor and method for measuring blood impedance phase angle |
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| CN111239228A true CN111239228A (en) | 2020-06-05 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112285171A (en) * | 2020-11-25 | 2021-01-29 | 三诺生物传感股份有限公司 | Impedance testing method applied to electrochemical test strip and electrochemical measuring system |
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