CN110487865A - A kind of production method of urinary albumin test electrode - Google Patents
A kind of production method of urinary albumin test electrode Download PDFInfo
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- CN110487865A CN110487865A CN201910833338.8A CN201910833338A CN110487865A CN 110487865 A CN110487865 A CN 110487865A CN 201910833338 A CN201910833338 A CN 201910833338A CN 110487865 A CN110487865 A CN 110487865A
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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
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/301—Reference electrodes
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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
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/307—Disposable laminated or multilayered electrodes
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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
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/308—Electrodes, e.g. test electrodes; Half-cells at least partially made of carbon
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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
- G01N27/28—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
- G01N27/3275—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction
- G01N27/3278—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction involving nanosized elements, e.g. nanogaps or nanoparticles
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Abstract
The invention discloses a kind of production methods of urinary albumin test electrode, comprising the following steps: the preparation of pet substrate；Graphene working electrode, reference electrode, the production to electrode；It uses print process that polymer insulation paint is printed in except insertion end and reaction zone are with the pet substrate of exterior domain, forms insulating layer；By crosslinking agent and urinary albumin selective reaction object solution, graphene working electrode, silver/silver chlorate are spread evenly across as reference electrode, carbon as the sample reaction zone to electrode in the method for spin coating, it is dried in 39-42 DEG C, then one layer of PVC insulating film is covered in reaction zone and form injection port, obtain urinary albumin test electrode.Urinary albumin of the invention has the advantages of time-consuming short, work efficiency is high when testing electrode detection；In addition, can accomplish the content for detecting urinary albumin in due course, the difficulty that twenty-four-hour urine sample is acquired when prior art detection is avoided.
Description
Technical field
The present invention relates to clinical medicines to examine field, and in particular to a kind of production method of urinary albumin test electrode.
Background technique
Urinary albumin is nephrosis early stage important sensitive indicator.It is also the angiosises such as kidney caused by diabetes, hypertension
Degenerative disease non-invasive monitoring goldstandard.Have become a great public health problem of China or even the whole world in recent decades,
The disease incidence of chronic kidney disease is worldwide in growth trend, 1.3 hundred million people of Patients with Chronic Kidney Disease in China year by year, be after
The another major disease for directly threatening human health after cardiovascular and cerebrovascular disease, diabetes, tumour.
But during present clinical medicine is examined, urinary albumin is examined and mostly uses enzyme-linked immunization, radioimmunology, efficiently greatly
Liquid chromatography and immunoturbidimetry.Wherein:
Enzyme-linked immunization detection time is longer；Radioimmunology sensitivity, accuracy are higher, but there are radioactivity dirts
Dye.
High performance liquid chromatography complex disposal process, operating process are cumbersome.
Immune scatter turbidimetry is the method that current clinical examination commonly measures urinary albumin, due to urinary albumin content
Very low (normal value is less than 20mg/L), the sensibility and specificity of Immune scatter turbidimetry can not all measure the accurate of urinary albumin
Value, therefore testing result cannot function as the foundation of Accurate Diagnosis kidney injury, and checkout procedure takes a long time, from patient's specimen taken
It is needed 4~5 hours to inspection result is obtained about, is unable to satisfy modernization precisely the easy, quick of medical treatment, Accurate Diagnosis and monitoring disease
The requirement of disease.
Summary of the invention
In order to realize rapid, the accuracy, high sensitivity of urinary albumin detection, it is an object of the invention to: provide one
The production method of kind urinary albumin test electrode.
To achieve the above object, the invention provides the following technical scheme:
A kind of production method of urinary albumin test electrode, comprising the following steps:
(1) it the preparation of pet substrate: directly buys；
(2) production of graphene working electrode:
(2.1) graphene working electrode is by serotonin and hydrophilic imidazole ion liquid IL 70-80 under hydrothermal conditions
DEG C isothermal reaction 7-9h, reduction-oxidation type two-dimensional graphene lamella take mutually product with lamella and connects recombination, obtain macroscopical three
Porous graphene assembly is tieed up, water is then dissolved in and obtains three-dimensional porous graphene assembling liquid solution；
(2.2) it disperses crosslinking agent in three-dimensional porous graphene assembling liquid solution with inorganic nano array material and obtains
Mixed solution；
(2.3) mixed solution is printed on pet substrate, forms graphene working electrode in 39-42 DEG C of drying；
(5) production of reference electrode:
Reference electrode is silver/silver chloride electrode, the production method is as follows: PET will be printed in after silver/silver chloride solution dissolution
Substrate surface；
(6) to the production of electrode:
It is carbon electrode to electrode, the production method is as follows: pet substrate surface is printed in after carbon solution is dissolved；
(5) production of insulating layer
It uses print process that polymer insulation paint is printed in except insertion end and reaction zone are with the pet substrate of exterior domain, is formed
Insulating layer, wherein the graphene working electrode, reference electrode are insertion end to the region where electrode leading portion, described
Graphene working electrode, reference electrode are reaction zone to the region where electrode back segment；
(6) production of urinary albumin test electrode:
By crosslinking agent and urinary albumin selective reaction object solution, graphene work electricity is spread evenly across in the method for spin coating
Pole, silver/silver chlorate, as the sample reaction zone to electrode, are dried, then in reaction zone as reference electrode, carbon in 39-42 DEG C
One layer of PVC insulating film is covered, forms injection port at the edge port of PVC insulating film, obtains urinary albumin test electrode.
Preferably, water, oxidized form two-dimensional graphene nanometer sheet, serotonin and hydrophilic imidazol ion in step (2.1)
The parts by weight of liquid IL are respectively 13-17 parts, 15-25 parts, 7-18 parts, 45-55 parts.
Preferably, crosslinking agent is polyaziridine in step (2.2), inorganic nano array material is carboxylic carbon nano-tube；
Wherein, polyaziridine, carboxylic carbon nano-tube, three-dimensional porous graphene assembling liquid solution parts by weight be respectively 2-4 parts,
20-35 parts, 45-60 parts.
Preferably, the length and width of pet substrate, thick respectively 28-30mm, 5.8-6.2mm, 0.7-0.9mm in step (1).
Preferably, the length and width of graphene working electrode, thick respectively 18-22mm, 5.8-6.2mm, 0.3-0.5mm；Reference
The length and width of electrode, thick respectively 18-22mm, 5.8-6.2mm, 0.3-0.5mm；Length and width, thick respectively 18- to electrode
22mm、5.8-6.2mm、0.3-0.5mm。
Preferably, the length and width of insulating layer, thick respectively 18-22mm, 5.8-6.2mm, 0.3-0.5mm in step (5).
Preferably, crosslinking agent is polyaziridine in step (6), and urinary albumin selective reaction object is brilliant cresyl blue；Water,
Polyaziridine, brilliant cresyl blue parts by weight be respectively 85-95 parts, 2-4 parts, 5-15 parts.
Preferably, the length and width of PVC insulating film, thick respectively 5-6mm, 1.5-2.5mm, 0.05-0.15mm in step (6).
Compared with prior art, the beneficial effects of the present invention are:
1) its electric conductivity is fabulous by using graphene working electrode by the present invention, and the electric current generated when detection is made almost
Reach zero loss, ensure that the accuracy of detection；2) reaction zone of urinary albumin test electrode of the present invention is selected by urinary albumin
Selecting property reactant (brilliant cresyl blue) can be good at avoiding the interference of other visible components in urine, and selective reaction object
(brilliant cresyl blue) reaction sensitivity is very high, is capable of detecting when the urinary albumin of extremely low concentration, substantially increases low concentration urine
The accuracy of albumin；It 3) is (magnificent by urinary albumin selective reaction object when urinary albumin test electrode detection of the present invention
Cresyl blue) curent change is generated with albumin reaction in urine obtains testing result, therefore, have time-consuming short, substantially increases
Working efficiency；In addition, can accomplish the content for detecting urinary albumin in due course, twenty-four-hour urine is acquired when avoiding prior art detection
The difficulty of sample.
Detailed description of the invention
Fig. 1 is the structural schematic diagram that the urinary albumin that better embodiment of the present invention provides tests electrode.
Fig. 2 is the side structure schematic diagram that urinary albumin of the present invention tests electrode.
In appended drawing reference: 10-PET substrate, tri- electrode of 20-, 21- graphene working electrode, 22- reference electrode, 23- are to electricity
Pole, 30- insulating layer, 40-PVC insulating film, 41- injection port, 50- reaction zone.
Specific embodiment
In order to make the objectives, technical solutions, and advantages of the present invention clearer, with reference to the accompanying drawings and embodiments, right
The present invention is further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, and
It is not used in the restriction present invention.
The production method of urinary albumin test electrode of the present invention, comprising the following steps:
1, the preparation of pet substrate: pet substrate is outsourcing part, and preferred size is 30mm x 6mm x0.8mm.
2, the production of three electrodes: three electrodes are respectively working electrode, reference electrode and the production to electrode.
1) production of working electrode: the graphene working electrode is by reducing agent serotonin (parts by weight 10%) and hydrophilic
Imidazole ion liquid IL (parts by weight 50%) (1-Butyl-3-methylimidazolium tetrafluoroborate,
BMIMBF4) 75 DEG C isothermal reaction 8 hours under hydrothermal conditions, reduction-oxidation type two-dimensional graphene (parts by weight 20%) lamella with
Lamella takes mutually product and connection recombination, and it is porous to construct macroscopic three dimensional using oxidized form two-dimensional graphene nanometer sheet as structural unit
Graphene assembly, then by the way that by crosslinking agent (polyaziridine) (parts by weight 3%), (carboxylated carbon is received with inorganic nano array material
Mitron) (parts by weight 30%) be scattered in three-dimensional porous graphene (parts by weight 60%) assembling liquid solution, by nano material and
Synergistic effect between graphene increases electrode active surface site and electrode adsorption ability, promotes between electrode and analyte
Electron transmission, the graphene assembling liquid solution for accelerating electrochemical reaction speed, the electrocatalysis characteristic for improving electrode；Then using print
Brush method is by three-dimensional porous graphene assembly, crosslinking agent (polyaziridine) and inorganic nano array material (carboxylated carbon nanometer
Pipe) mixed solution is printed in pet substrate, graphene working electrode is formed in 40 DEG C of drying.
2) production of reference electrode: reference electrode is silver/silver chloride electrode, and silver/silver chlorate is that commercially available analysis is pure.After dissolution
It is printed in pet substrate surface.It is preferred that printed dimensions are 20mm x 0.9mm x 0.3mm.
3) to the production of electrode: being carbon electrode to electrode.Carbon is commercially available activated carbon, and pet substrate surface is printed in after dissolution.
It is preferred that printed dimensions are 20mm x 0.9mm x 0.3mm.
3, use print process that polymer insulation paint is printed in except insertion end and reaction zone are with the pet substrate of exterior domain, shape
At insulating film, insulated paint is commercially available.It is preferred that printed dimensions are 20mm x 6mm x 0.4mm.
4, the rhetorical functionization in electrode reaction area is made urinary albumin and tests electrode: crosslinking agent (polyaziridine) (parts by weight
3%) it with urinary albumin selective reaction object (brilliant cresyl blue) solution (parts by weight 10%), is spread evenly across in the method for spin coating
Graphene working electrode, silver/silver chlorate, as the sample conversion zone to electrode, are dried, so as reference electrode, carbon in 40 DEG C
One layer of PVC insulating layer cover film is covered in reaction zone afterwards and forms injection port, obtains urinary albumin test electrode.PVC insulating layer
Cover film preferred size is 5.5mm x 2.0mm x 0.1mm.
Urinary albumin of the present invention test electrode is that current medical test circle in Analysis of urine albumin first enters electrochemical test
Research field.
Principle of products: the electrode is inserted on Electrochemical Detection platform, platform gives electrode one constant voltage, makes electricity
Stream flows through working electrode and constitutes a system that is obstructed or being powered less substantially to electrode, working electrode and reference electrode, utilizes
The stability of reference electrode current potential measures the electrode potential of working electrode.Working electrode and the body that an energization is constituted to electrode
System, for measuring the electric current that working electrode passes through.By urine to be measured close to the well of electrode, electrode is automatically complete by siphon
Electrode reaction area, the urinary albumin selective substances brilliant cresyl blue and urine of reaction zone are tested at quantitative suction sample to urinary albumin
In albumin reacted, so as to cause the variation of electric current in original working electrode and the system being powered to electrode, and electric current
The size of variation is directly proportional to albumin content in urine, reads curent change size by electrochemistry platform, preceding with test
The current curve figure for knowing that the urinary albumin calibration object of concentration is drawn compares automatically, calculates urinary albumin content.
The present invention realizes clinical examination completion detection quick when urinary albumin is examined, in due course, accurate, is that other show
There is detection method incomparable.
Experimental result:
6 batches are taken to be purchased from the microdose urine protein quality-control product of the different known concentrations of Britain's Landau production, respectively with exempting from
Epidemic disease turbidimetry and urinary albumin of the invention test electrode are tested.
Instrument: Shanghai Chen Hua CHI600E electrochemical analyser, 7600 Biochemical Analyzer of Hitachi
Reagent: graceful microdose urine protein assay kit (immunoturbidimetry) lot number in Neusoft Witter
MLB1807019
Method: 6 batch microdose urine protein quality-control products are tested into electrode with urinary albumin provided by the invention respectively and are existed
The graceful microdose urine protein assay kit (immunoturbidimetry) of Shanghai Chen Hua CHI600E electrochemical analyser and Neusoft Witter is in day
It is tested on vertical 7600 Biochemical Analyzers, each batch is tested 3 times respectively.
Conclusion: the very close Quality Control target value of test result of urinary albumin test electrode of the present invention, and immunoturbidimetry
Test result deviates Quality Control target value.Above experiments have shown that urinary albumin test electrode of the invention can accurately detect urinary albumin
Content.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all in essence of the invention
Made any modifications, equivalent replacements, and improvements etc., should all be included in the protection scope of the present invention within mind and principle.
Claims (8)
1. a kind of production method of urinary albumin test electrode, it is characterised in that: the following steps are included:
(1) it the preparation of pet substrate: directly buys；
(2) production of graphene working electrode:
(2.1) graphene working electrode is by serotonin and hydrophilic imidazole ion liquid IL 70-80 DEG C of perseverance under hydrothermal conditions
Temperature reaction 7-9h, reduction-oxidation type two-dimensional graphene lamella take mutually product with lamella and connect recombination, and the three-dimensional for obtaining macroscopic view is more
The graphene assembly in hole is then dissolved in water and obtains three-dimensional porous graphene assembling liquid solution；
(2.2) it disperses crosslinking agent in three-dimensional porous graphene assembling liquid solution with inorganic nano array material and is mixed
Solution；
(2.3) mixed solution is printed on pet substrate, forms graphene working electrode in 39-42 DEG C of drying；
(3) production of reference electrode:
Reference electrode is silver/silver chloride electrode, the production method is as follows: pet substrate will be printed in after silver/silver chloride solution dissolution
Surface；
(4) to the production of electrode:
It is carbon electrode to electrode, the production method is as follows: pet substrate surface is printed in after carbon solution is dissolved；
(5) production of insulating layer
It uses print process that polymer insulation paint is printed in except insertion end and reaction zone are with the pet substrate of exterior domain, forms insulation
Layer, wherein the graphene working electrode, reference electrode are insertion end, the graphite to the region where electrode leading portion
Alkene working electrode, reference electrode are reaction zone to the region where electrode back segment；
(6) production of graphene brilliant cresyl blue urinary albumin test electrode:
By crosslinking agent and urinary albumin selective reaction object solution, with the method for spin coating be spread evenly across graphene working electrode,
As reference electrode, carbon as the sample reaction zone to electrode, then Yu Yu 39-42 DEG C drying adds silver/silver chlorate in reaction zone
One layer of PVC insulating film of lid forms injection port at the edge port of PVC insulating film, obtains graphene brilliant cresyl blue and urinates white egg
White test electrode.
2. the production method of urinary albumin according to claim 1 test electrode, it is characterised in that: water in step (2.1),
The parts by weight of oxidized form two-dimensional graphene nanometer sheet, serotonin and hydrophilic imidazole ion liquid IL are respectively 13-17 parts,
15-25 parts, 7-18 parts, 45-55 parts.
3. the production method of urinary albumin test electrode according to claim 1, it is characterised in that: handed in step (2.2)
Connection agent is polyaziridine, inorganic nano array material is carboxylic carbon nano-tube；Wherein, polyaziridine, carboxylic carbon nano-tube,
The parts by weight of three-dimensional porous graphene assembling liquid solution are respectively 2-4 parts, 20-35 parts, 45-60 parts.
4. the production method of urinary albumin test electrode according to claim 1, it is characterised in that: PET base in step (1)
The length and width of plate, thick respectively 28-30mm, 5.8-6.2mm, 0.7-0.9mm.
5. the production method of urinary albumin test electrode according to claim 1, it is characterised in that: graphene working electrode
Length and width, it is thick be respectively 18-22mm, 5.8-6.2mm, 0.3-0.5mm；The length and width of reference electrode, it is thick be respectively 18-22mm,
5.8-6.2mm,0.3-0.5mm；Length and width, thick respectively 18-22mm, 5.8-6.2mm, 0.3-0.5mm to electrode.
6. the production method of urinary albumin test electrode according to claim 1, it is characterised in that: insulation in step (5)
The length and width of layer, thick respectively 18-22mm, 5.8-6.2mm, 0.3-0.5mm.
7. the production method of urinary albumin test electrode according to claim 1, it is characterised in that: crosslinking in step (6)
Agent is polyaziridine, and urinary albumin selective reaction object is brilliant cresyl blue；The parts by weight of water, polyaziridine, brilliant cresyl blue
Respectively 85-95 parts, 2-4 parts, 5-15 parts.
8. the production method of urinary albumin test electrode according to claim 1, it is characterised in that: PVC is exhausted in step (6)
The length and width of velum, thick respectively 5-6mm, 1.5-2.5mm, 0.05-0.15mm.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111044585A (en) * | 2019-12-31 | 2020-04-21 | 浙江大学 | Double-test-strip levodopa detection system and method thereof |
| TWI803992B (en) * | 2020-09-28 | 2023-06-01 | 中央研究院 | Biosensor, bio-sensing system comprising the same and method for preparing the same |
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| EP1225449A1 (en) * | 2001-01-19 | 2002-07-24 | Apex Biotechnology Corporation | Non-enzymatic disposable electrode strip comprising a surfactant for detecting uric acid or hemoglobin; method for producing the same and its use |
| CN103954673A (en) * | 2014-03-25 | 2014-07-30 | 西北师范大学 | Method for applying ionic liquid functionalized graphene modified electrode in detection of 5-hydroxytryptamine and dopamine |
| CN106814123A (en) * | 2016-12-19 | 2017-06-09 | 华中科技大学 | A kind of ion liquid functionalization graphene paper electrode of Jenner's popped rice modification, its preparation method and application |
| CN109444234A (en) * | 2018-10-23 | 2019-03-08 | 安徽信灵检验医学科技有限公司 | A kind of production method of graphene brilliant cresyl blue urinary albumin test electrode |
| CN109776830A (en) * | 2019-03-05 | 2019-05-21 | 中原工学院 | A kind of polyurethane/carboxymethyl chitosan/PAn conducting membrane preparation method |
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2019
- 2019-09-04 CN CN201910833338.8A patent/CN110487865A/en active Pending
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| Publication number | Priority date | Publication date | Assignee | Title |
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| EP1225449A1 (en) * | 2001-01-19 | 2002-07-24 | Apex Biotechnology Corporation | Non-enzymatic disposable electrode strip comprising a surfactant for detecting uric acid or hemoglobin; method for producing the same and its use |
| CN103954673A (en) * | 2014-03-25 | 2014-07-30 | 西北师范大学 | Method for applying ionic liquid functionalized graphene modified electrode in detection of 5-hydroxytryptamine and dopamine |
| CN106814123A (en) * | 2016-12-19 | 2017-06-09 | 华中科技大学 | A kind of ion liquid functionalization graphene paper electrode of Jenner's popped rice modification, its preparation method and application |
| CN109444234A (en) * | 2018-10-23 | 2019-03-08 | 安徽信灵检验医学科技有限公司 | A kind of production method of graphene brilliant cresyl blue urinary albumin test electrode |
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| CN111044585A (en) * | 2019-12-31 | 2020-04-21 | 浙江大学 | Double-test-strip levodopa detection system and method thereof |
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