CN106124595A - A kind of biosensor and preparation method thereof and reaction reagent - Google Patents
A kind of biosensor and preparation method thereof and reaction reagent Download PDFInfo
- Publication number
- CN106124595A CN106124595A CN201610436485.8A CN201610436485A CN106124595A CN 106124595 A CN106124595 A CN 106124595A CN 201610436485 A CN201610436485 A CN 201610436485A CN 106124595 A CN106124595 A CN 106124595A
- Authority
- CN
- China
- Prior art keywords
- reaction reagent
- biosensor
- electrode
- layer
- reagent
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- 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
Abstract
The present invention relates to biological detection instrument field, it is specifically related to a kind of biosensor improving stability, its composition includes insulated substrate, the electrode system being attached on insulated substrate, described electrode system at least includes that a working electrode and one are to electrode, cover the reaction reagent layer on electrode system, described reaction reagent layer should at least cover a working electrode, and it is positioned at the reative cell above electrode system and reaction reagent layer, described reative cell is made up of middle interlayer and cover layer, and reative cell has air-vent, wherein: containing one or more in imidazoles or imidazole derivative in the reaction reagent that described reaction reagent layer uses.The present invention significantly improves the problem of the background current rising that biosensor increases with the increase of storage time, thus improves the storage stability of biosensor, it is ensured that its test accuracy before the deadline.
Description
Technical field
The present invention relates to biological detection instrument field, be specifically related to a kind of biosensor improving stability and preparation thereof
Method and reaction reagent.
Background technology
Biosensor (biosensor), is that one utilizes bioactive substance Selective recognition target substance and by it
Concentration or content are converted to the instrument that the signal of telecommunication carries out detecting.It is to be made recognition component by immobilized biological sensitive materials (to include
The bioactive substances such as enzyme, antibody, antigen, microorganism, cell, tissue, nucleic acid), suitable physics and chemistry transducer (as oxygen electrode,
Photosensitive tube, field effect transistor, piezoquartz etc.) and signal amplifying apparatus constitute analytical tool or system.
Electrochemica biological sensor has been widely used in in-vitro diagnosis field, and these instrument systems can detect in body fluid
Physiologically substance, facilitate doctor to patient status analysis and diagnosis, also have some instrument systems to be applied to family and use, convenient disease
The self-monitoring of people, these physical signs include blood glucose, cholesterol, uric acid, triglyceride, lactic acid, ketoboidies, enzyme etc..
Biosensor has been widely used in Clinical detection at present, and has some wherein biosensor products
Being widely popularized to the daily monitoring of domestic, become universal daily consumable goods, most popular example is that portable blood sugar is surveyed
Test system.For the use of biosensor, especially domestic, especially due to storage condition condition and the limit of use frequency
System, it is desirable to biosensor product has looser storage environmental requirement and longer effect duration.And for storage condition bar
Part and the restriction of effect duration, its topmost factor is the stability of reaction reagent.
Reaction reagent generally includes enzyme (also having some without enzyme sensor), electron transfer mediator, polymer, buffer, table
Face activating agent, stabilizer etc., wherein affect stability it is crucial that enzyme and electron transfer mediator, it usually needs adds stabilizer and changes
Its stability kind.Either enzyme or electron transfer mediator, under higher temperature and humidity, all can show instability, enzyme
Meeting loss of activity gradually, electron transfer mediator, it may happen that reaction, causes the deviation of test result, limits the use of product
The effect phase.Biosensor product is commonly stored in sealing drum or the bag of band desiccant at present, keeps the drying property of environment, to change
Kind stability.But storing containers is not positive confinement, and repeat switch taken by container, and dampness also can progress into,
Adding heat is also to be relatively difficult to isolation, the therefore stability of a reaction reagent difficult problem the most in the industry.
Modal electron transfer mediator is the iron cyanide, has the feature that cost is cheap and reaction suitability is good, application
Widely.But the iron cyanide is under higher temperature and humidity, including the water retained in the dampness in container, reaction reagent
Point, hydrophilic substance in reagent, it may occur that oneself or the redox reaction intersected, produce and detect reaction similar in the middle of produce
Thing, ultimately results in the increase of background signal, makes the false rising of detection reaction signal, and the detection having a strong impact on biosensor is accurate
Degree.
The United States Patent (USP) that open (bulletin) number is US6911131B2, open (bulletin) day is on 06 28th, 2005, discloses
A kind of biosensor improving storage stability, its core content is interpolation sugar alcohol, slaine, molecule in reaction reagent
It is contained within the organic acid of at least one carboxyl or acylate or intramolecular containing at least one carboxyl and the organic acid of amino or to have
Any one or combination in machine hydrochlorate, and therefore suppress to increase with the storage time and the rising of background current that increases.But,
Inventor is found by research, and the method still exists the phenomenon that obvious background current raises with the storage time.And for example, open
The United States Patent (USP) that (bulletin) number is US20130026050A1, open (bulletin) day is on 01 31st, 2013, discloses improvement ferrum
The another kind of method of cyanide performance, its core content is the cationic salts filtering out the suitable iron cyanide, and this patent discloses
Cation be tetramethyl-ammonium, and therefore suppress the rising of background current increased because the iron cyanide increases with the storage time.
But tetramethyl-ammonium is poisonous, heating is easily decomposed, and the course of processing is impracticable, and inventor is found by research, and the method also depends on
So there is the phenomenon that obvious background current raises with the storage time.
Summary of the invention
For overcoming the problem and defect that above-mentioned prior art exists, the purpose of the present invention aims to provide one to be had and well deposits
The biosensor of storage stability, it is ensured that its test accuracy within the effect phase.
For realizing the purpose of the present invention, inventor provides following technical scheme:
Inventor provide firstly a kind of biosensor, and its composition includes insulated substrate, is attached on insulated substrate
Electrode system, described electrode system at least includes that a working electrode and one, to electrode, cover on electrode system
Reaction reagent layer, described reaction reagent layer should at least cover a working electrode, and be positioned at electrode system and reaction reagent
Layer reative cell above, described reative cell is made up of middle interlayer and cover layer, and reative cell has air-vent, wherein: described
The reaction reagent that uses of reaction reagent layer in containing one or more in imidazoles or imidazole derivative.
Present invention is generally directed to is the stability improving electron transfer mediator, and the core content of the present invention is reaction reagent
Composition.Research finds, imidazoles or imidazole derivative can effectively suppress the redox reaction that electron transfer mediator is potential, from
And significantly improve the problem of the background current rising that biosensor increases with the increase of storage time, thus promote bio-sensing
The storage stability of device, it is ensured that its test accuracy before the deadline.The composition of reaction reagent include electron transfer mediator and
Stabilizer, described electron transfer mediator is that the iron cyanide, imidazoles or imidazole derivative can add as stabilizer additive；Miaow
Azoles or imidazole derivative also can add as buffer.Add preferably as buffer.As current techique, reaction examination
Agent, according to the difference of detection target, can include specific reaction enzymes；Reaction reagent as required, can add suitable surface
Other components such as activating agent, polymer, buffer, repeat the most one by one.
In the present invention, the material of described insulated substrate includes but not limited to the one in PET, PP, PVC, acrylic.
Described electrode system, builds on insulated substrate, and electrode system at least includes working electrode and to electrode；Each
Electrode has extension wire to extend to the other end of substrate, is formed and connects haptic element；Extension wire, connection haptic element and reaction electrode
Material can be identical, it is also possible to different.After biosensor testing plate inserting instrument, test sheet connects by connecting haptic element and instrument
The connection connecing device realizes both signal communications.The material of electrode includes but not limited to graphite, silver, gold, platinum, palladium, ITO and IZO
In one.
Described reaction reagent is fixed on electrode system, at least covers working electrode.
Described middle interlayer builds above electrode system, and reative cell region hollow out defines reative cell in conjunction with cover layer
I.e. sample intake passage, middle interlayer material can be band base material or the adhesive tape without base material, bonds up after processing；Can also be glue
Or polymer paste, got on by silk screen printing；As separated without other insulant on electrode, in every material must be good
Insulant.
Described cover layer is above middle interlayer, and middle interlayer, substrate are built into reative cell i.e. sample intake passage, in order to pass through
Siphonage auto injection.Cover layer can use PET material, the material that the hydrophilic treated of preferably clear is crossed, and transparent can help is used
Family easily confirms reaction zone sample introduction state, hydrophilic sample introduction can be made more smoothly, and non-reative cell region can be opaque, prints sample introduction
Mark and Logo, 9971 hydrophilic film of available material such as 3M company.
Described reative cell should have air-vent, in order to formed siphonage, air-vent can build insulated substrate or in
On interlayer or cover layer.
As preferably, in the present invention, described imidazoles or the imidazole derivative addition in reaction reagent be 1mM～
2M.More preferably 20mM～200mM.
As preferably, in the present invention, described imidazole derivative includes being not limited to following: 1-Methylimidazole., 1-second
Base imidazoles, 1-propyl imidazole, 2,4-methylimidazole, 2-methylimidazole, 2-ethyl imidazol(e), 4-methylimidazole, 1,2-dimethyl
Imidazoles, 2-ethyl-4-methylimidazole and 1-ethoxy-2-methylimidazole etc..
As preferably, in the present invention, described reaction reagent includes being not limited to following kind: glucose response reagent,
Lentochol reaction reagent, uric acid reaction reagent, ketoboidies reaction reagent, lactic acid reaction reagent and hemoglobin reaction reagent etc..
As preferably, in the present invention, the thickness of described middle interlayer is 0.025mm～0.5mm.As it is further preferred that in every
The thickness of layer is 0.075mm～0.125mm.
Present invention also offers the preparation method of above-mentioned a kind of biosensor, including:
(1) forming electrode system on insulated substrate, described electrode system at least includes a working electrode and one
To electrode, processing method has: by silk screen printing, specific electrode pattern is printed onto on insulated substrate, then by heating or
Infrared or ultraviolet is dried；Or electrode material is attached on insulated substrate by the way of sputtering, then by laser ablation
Mode processes specific electrode pattern,
(2) reaction reagent layer is fixed to electrode by the processing of a liquid, silk screen printing, inkjet printing or crack extrusion method
On,
(3) fit on electrode system and reaction reagent layer the middle interlayer of reative cell and cover layer, complete, wherein:
Adhesive tape with band base material or without base material is middle interlayer material, is bonded in electrode system and reaction reagent layer after processing；Or
Person is middle interlayer material with glue or polymer paste, by being screen printed onto in electrode system and reaction reagent layer.
The present invention has also carried the reaction reagent of a kind of biosensor, spreads out containing imidazoles or imidazoles in described reaction reagent
One or more in biology.
The core content of the present invention is the composition of reaction reagent, is primarily directed to improve stablizing of electron transfer mediator
Property.Research finds, imidazoles or imidazole derivative can effectively suppress the redox reaction that electron transfer mediator is potential, thus aobvious
Write the problem improving the background current rising that biosensor increases with the increase of storage time, thus promote biosensor
Storage stability, it is ensured that its test accuracy before the deadline.
The composition of reaction reagent generally includes enzyme (also having some without enzyme sensor), electron transfer mediator, polymer, buffering
Liquid, surfactant and stabilizer etc..The effect of enzyme is specific and detected material reacts；Electron transfer mediator is from enzyme and quilt
Obtaining electronics in the reaction of detectable substance, and be diffused on electrode release electronics, and form signal, this signal can be caught with tested instrument
Obtain；Buffer ensures a suitable reaction environment；Surfactant can optimize reaction condition, and promotes that sample spreads；Many
Polymers can form grid structure, and immobilized enzyme to a certain extent.
Different according to detection purpose, different enzymes can be selected, as detection blood glucose can select glucose dehydrogenase, Portugal
Glucoseoxidase；Detection lactic acid, can select Lactate Oxidase, lactic acid dehydrogenase；Detection uric acid, can select uric acid to aoxidize
Enzyme, it is also possible to select the acid sensor without enzymuria without enzyme；Detection beta-hydroxybutyric acid, it is possible to use beta-hydroxybutyric dehydrogenase etc.；
Separately some needs the parameter of two or more enzyme, such as cholesterol detection, needs cholesteryl esterase and cholesterol oxidase or gallbladder solid
The combination of alcoholdehydrogenase；The combination of some oxidase and peroxidase carries out detection etc..
Representational electron transfer mediator has ferrocene derivatives, such as ferrocenecarboxylic acid；The iron cyanide, such as the potassium ferricyanide；
Ruthenium complex, such as six ammonia ruthenium trichlorides；Prussian blue；Methyl blue；Mai Er Doran；Toluphenazine Methylsulfate salt compounds；
2,6 dichlorophenol indophenol；Quinones；Cobalt, nickel complex；Four nitre azole compounds；Naphthols chlorine；4-AA etc..
The iron cyanide is cheap because of its cost, and water solublity and reaction suitability are good, and application widely, present invention is generally directed to this electronics and passes
Research made by the biosensor passing amboceptor.
Polymer includes binding agent and filler etc..Binding agent can be xanthan gum, gelatin, hydroxyethyl cellulose, hydroxyl first
Base propyl cellulose etc.；Filler can be cellulose family, such as microcrystalline Cellulose, methylcellulose, ethyl cellulose, ethoxy
Cellulose, hydroxymethyl-propyl cellulose, sodium carboxymethyl cellulose, carboxymethyl starch sodium, apatite, gasification silicon dioxide, kaolinite
Soil, montmorillonite, chitosan, cyclodextrin, sodium alginate, glucosides, PMMA, agar, gelatin, Polyethylene Glycol, polyvinylpyrrolidone,
PEO, PVA, PVP/VA etc..
Alternative surfactant has: Triton X-100 class, such as Triton X-100；Polyoxyethylene loses
Water sorbitan monolaurate, such as Tween 20；Chlolic acid derivatives class, as 3-[(3-gallbladder amidopropyl) dimethylammonio]-
1-propane sulfonic acid salt, NaTDC；Brij-35, such as Brij 35；Quaternary ammonium salt, such as cetyl trimethyl
Ammonium bromide；Glucosides class, such as dodecyl-β-D-Maltose glycosides；Poly-(oxo-1,2-second dimethylene)-α-nonyl phenyl-ω-
Hydroxy kind, such as TERGITOL (TM) NP-40, etc..
Alternative buffer is also a lot, owing to activity and the stable region of enzyme limit, and the pH value of the buffer of selection
Scope is generally between 3～11.Typical buffer has: acetate buffer, citrate buffer solution, phosphate buffer,
TES, BES, HEPES, PIPES, MOPS, Bicine, Tris, CHES, MES, ADA, CAPS, EPPS, POPSO, CAPS etc..
Each component of above-mentioned reaction reagent can be combined according to the needs of detection material, and electron transfer mediator is preferential
Select that cost is cheap, good water solubility, the iron cyanide that reaction suitability is good.The core of the present invention is that reaction reagent includes stable
Agent, described stabilizer is the one or more combination of imidazoles or imidazole derivative；Imidazoles or imidazole derivative also can conducts
Buffer adds reaction reagent.Add preferably as buffer, this is because usual reagent needs comprise buffer control pH
Value is to reach optimal reaction environment, but such as adds other buffer, and its buffer composition may result in electron mediator not
Stable；And imidazoles or imidazole derivative add as buffer, play buffer and the effect of stabilizer simultaneously, can reduce
Add the stability risk that other buffer cause, the cost adding other buffer can also be saved simultaneously.The one of the present invention
Individual embodiment also demonstrates this viewpoint.
As preferably, in the reaction reagent of the present invention, described imidazoles or imidazole derivative making in reaction reagent
It is 1mM～2M with concentration ranges.More preferably 20mM～200mM.
As preferably, in the reaction reagent of the present invention, described imidazole derivative includes being not limited to following: 1-methyl
Imidazoles, 1-ethyl imidazol(e), 1-propyl imidazole, 2,4-methylimidazole, 2-methylimidazole, 2-ethyl imidazol(e), 4-methylimidazole, 1,
2-methylimidazole, 2-ethyl-4-methylimidazole and 1-ethoxy-2-methylimidazole etc..
As preferably, described reaction reagent kind includes being not limited to: glucose response reagent, lentochol reaction try
Agent, uric acid reaction reagent, ketoboidies reaction reagent, lactic acid reaction reagent and hemoglobin reaction reagent etc..
Compared with prior art, the invention have the advantages that
The present invention introduces imidazoles or imidazole derivative in reaction reagent and adds as stabilizer or buffer addition.Miaow
Azoles or imidazole derivative can effectively suppress the redox reaction that electron transfer mediator is potential, thus significantly improve bio-sensing
The problem that the background current that device increases with the storage time and increases raises, thus promote the storage stability of biosensor, protect
Demonstrate,prove its test accuracy before the deadline.
Experimental results shows, the reagent controls of the present invention does not add the reaction of imidazoles or imidazole derivative
Reagent at 65 DEG C 2 weeks accelerated ageing rear backdrop electric current (0mg/dL) compare room temperature background current elevation amplitude and substantially to lack,
Illustrate that the present invention is effectual in terms of improving the stability of biosensor.
Accompanying drawing explanation
Figure 1A and Figure 1B is STRUCTURE DECOMPOSITION schematic diagram and the structural integrity schematic diagram of a kind of biosensor of the present invention.
Fig. 2 A-2D be the blood sugar test sheet accelerated ageing test of reaction reagent A～D of the present invention be 0mg/dL in concentration of specimens
Under background current curve chart.
Fig. 3 is blood sugar test sheet accelerated ageing and the post of 5 seconds background currents of room temperature control of reaction reagent A～D of the present invention
Shape comparison chart.
Detailed description of the invention
Below in conjunction with embodiment, further illustrate present disclosure.Should be appreciated that the enforcement of the present invention is not limited to
In the following examples, any pro forma accommodation and/or the change of being made the present invention fall within scope.
In the present invention, if not refering in particular to, all of equipment and raw material etc. are all commercially available or the industry is conventional.
Method in following embodiment, if no special instructions, is the conventional method of this area.
Embodiment 1
As shown in FIG. 1A and 1B, a kind of biosensor, its composition includes insulated substrate, and the material of insulated substrate is
PET, the electrode system being attached on insulated substrate, described electrode system includes that a working electrode and one are to electrode, electricity
Pole material is graphite, and each electrode has extension wire to extend to the other end of substrate, forms connection haptic element (during use, raw
After thing product sensor i.e. tests sheet inserting instrument, test sheet realizes both by connecting the connection of haptic element and instrument connector
Signal communication), cover the reaction reagent layer on electrode system, described reaction reagent layer covers working electrode, Yi Jiwei
Reative cell above electrode system and reaction reagent layer, described reative cell is made up of middle interlayer and cover layer, middle interlayer
Thickness is 0.125mm, and cover layer uses 9971 hydrophilic film of 3M company, and reative cell has air-vent, in order to forms siphon and makees
With, wherein: described reaction reagent layer uses reaction reagent B (imidazole buffer), and reaction reagent B is prepared as follows:
Take imidazoles 0.6808g, be dissolved in 80g water, with 1M salt acid for adjusting pH value to 6.2.Add following material, dissolve successively:
Gelatin (Sigma G7041): 0.0500g,
Sodium carboxymethyl cellulose (Sigma C5678): 1.0000g,
Polyethylene Glycol (Sigma 95904): 0.5000g,
Triton X-100:0.2000g,
The potassium ferricyanide: 3.2900g,
Glucoseoxidase (Amano, GO-AM): 600KU,
After all adding and be completely dissolved, add purified water constant volume to 100mL, standby.
Biosensor preparation method is as follows:
(1) by silk screen printing, specific electrode pattern is printed onto on insulated substrate, then heat drying, electrode system
It is attached on insulated substrate,
(2) reaction reagent B is assigned on the graphite electrode that silk screen printing makes by a liquid mode, then in temperature 55 DEG C
Lower drying 15 minutes, reaction reagent layer is fixing to be worked on electrode,
(3) fit on electrode system and reaction reagent layer the middle interlayer of reative cell and cover layer, complete, obtain one
Kind of blood sugar test sheet, wherein: be middle interlayer material with the adhesive tape of band base material, is bonded in electrode system and reaction reagent after processing
On layer.
Embodiment 2
The present embodiment structure and the course of processing are with embodiment 1, but reaction reagent is different, and described reaction reagent layer uses anti-
Answering reagent C (comparison+imidazoles), reaction reagent C is prepared as follows:
Disodium hydrogen phosphate,anhydrous 0.2626g, AMSP 0.9778g is added in 80g purified water；It is completely dissolved
Rear addition imidazoles 0.6808g, with using 1M salt acid for adjusting pH value to 6.2 after dissolving.And add following material, dissolve successively:
Gelatin (Sigma G7041): 0.0500g,
Sodium carboxymethyl cellulose (Sigma C5678): 1.0000g,
Polyethylene Glycol (Sigma 95904): 0.5000g,
Triton X-100:0.2000g,
The potassium ferricyanide: 3.2900g,
Glucoseoxidase (Amano, GO-AM): 600KU,
After all adding and be completely dissolved, add purified water constant volume to 100mL, standby.
Then according to embodiment 1 preparation method is fabricated to biosensor, obtain a kind of blood sugar test sheet.
Embodiment 3
The present embodiment structure and the course of processing are with embodiment 1, but reaction reagent is different, and described reaction reagent layer uses anti-
Answering reagent D (comparison+1-Methylimidazole .), reaction reagent D is prepared as follows:
Disodium hydrogen phosphate,anhydrous 0.2626g, AMSP 0.9778g is added in 80g purified water；It is completely dissolved
Rear addition 1-Methylimidazole. 0.8210g, with using 1M salt acid for adjusting pH value to 6.2 after dissolving.And add following material, the most molten
Solve:
Gelatin (Sigma G7041): 0.0500g,
Sodium carboxymethyl cellulose (Sigma C5678): 1.0000g,
Polyethylene Glycol (Sigma 95904): 0.5000g,
Triton X-100:0.2000g,
The potassium ferricyanide: 3.2900g,
Glucoseoxidase (Amano, GO-AM): 600KU,
After all adding and be completely dissolved, add purified water constant volume to 100mL, standby.
Then according to embodiment 1 preparation method is fabricated to biosensor, obtain a kind of blood sugar test sheet.
Comparative example
This comparative example structure and the course of processing are with embodiment 1, but reaction reagent is different, and described reaction reagent layer uses anti-
Answering reagent A (comparison), reaction reagent A is prepared as follows:
Purified water: 80g,
Disodium hydrogen phosphate,anhydrous: 0.2626g,
AMSP: 0.9778g,
Gelatin (Sigma G7041): 0.0500g,
Sodium carboxymethyl cellulose (Sigma C5678): 1.0000g,
Polyethylene Glycol (Sigma 95904): 0.5000g,
Triton X-100:0.2000g,
The potassium ferricyanide: 3.2900g,
Glucoseoxidase (Amano, GO-AM): 600KU,
Add above material, dissolve successively, wait after all adding and being completely dissolved, add purified water constant volume to 100mL, standby
With.
Then according to embodiment 1 preparation method is fabricated to biosensor, obtain a kind of blood sugar test sheet.
Experimental example
Four kinds of blood sugar test sheets embodiment 1-3 and comparative example being made detect.Every kind of blood sugar test sheet is divided into
Two parts, a part is stored in the sealed reagent cylinder of band desiccant and is placed on room temperature as test comparison；Another part stores
Accelerated ageing in 65 DEG C of baking ovens it is positioned over, test sheet after accelerated ageing in 65 DEG C of baking ovens in the sealed reagent cylinder of band desiccant
Be equivalent to storage at room temperature for a long time.After two weeks, the test sheet of accelerated ageing in 65 DEG C of baking ovens takes out, and is placed on the survey of room temperature
Test piece is tested together, altogether 3 concentration of test: 0mg/dL, 100mg/dL, 450mg/dL.Comparative example and the test result of embodiment
See Fig. 2 A, Fig. 2 B, Fig. 2 C, Fig. 2 D and Fig. 3 and following form:
The result of comparative example:
The result of embodiment 1:
The result of embodiment 2:
The result of embodiment 3:
Showing of above table, tests data from accelerated ageing, and three kinds are added imidazoles or the enforcement of imdazole derivatives
Example reaction reagent B, C, D compared to comparative example reaction reagent A, 2 weeks accelerated ageing rear backdrop electric current (0mg/ at temperature 65 DEG C
DL) comparing room temperature background current elevation amplitude substantially to lack, this point the most significantly embodies in 100mg/dL, 450mg/dL concentration
Out, wherein behave oneself best with embodiment 1 reaction reagent B.
Visible, reaction reagent adds imidazoles or imdazole derivatives and can effectively suppress the potential oxidation of electron transfer mediator also
Former reaction, thus significantly improve the problem of the background current rising that biosensor increases with the increase of storage time, thus carry
Rise the storage stability of biosensor, it is ensured that its test accuracy before the deadline.Wherein, as preferably, imidazoles or miaow
The derivant of azoles can be added as buffer.
Fig. 2 A～2D is the test sheet accelerated ageing test of reaction reagent A～D background under concentration of specimens is 0mg/dL
Current curve diagram, the curve of triangular data points is the curve at temperature 65 DEG C after accelerated ageing in 2 weeks, and square data points is for depositing
Store up in room temperature as the curve compareed.Visible three kinds are added imidazoles or the embodiment reaction reagent B of imdazole derivatives, and C, D compare
Reference examples reaction reagent A, the curve ratio room temperature control curve after 65 DEG C of accelerated ageings in 2 weeks raises and substantially to lack, and wherein B shows
Good.
Fig. 3 is blood sugar test sheet accelerated ageing and the cylindricality comparison of 5 seconds background currents of room temperature control of reaction reagent A～D
Figure.As seen from the figure, three kinds are added imidazoles or the embodiment reaction reagent B of imdazole derivatives, and C, D compare reference examples reaction reagent A,
After 65 DEG C of accelerated ageings in 2 weeks fewer than the rising of 5 seconds background currents of room temperature control, wherein B raise minimum.
Embodiment 4-12
Other are with embodiment 1, and difference is to replace with imidazoles the one in following material:
1-ethyl imidazol(e) (concentration in reaction reagent is 1mM), 1-propyl imidazole (concentration in reaction reagent is 5mM),
2,4-methylimidazole (concentration in reaction reagent is 20mM), 2-methylimidazole (concentration in reaction reagent is 100mM), 2-
Ethyl imidazol(e) (concentration in reaction reagent is 200mM), 4-methylimidazole (concentration in reaction reagent is 50mM), 1,2-diformazan
Base imidazoles (concentration in reaction reagent is 1M), 2-ethyl-4-methylimidazole and 1-ethoxy-2-methylimidazole (reaction reagent
In concentration be 2M).
Through test, at least can reach technique effect in the same manner as in Example 1, repeat no more.Particularly evident, implement
Example 6-9, when namely imidazole derivative concentration interval in reaction reagent is 20mM～200mM, imidazoles derives
Thing effectively suppresses the effect of the potential redox reaction of electron transfer mediator more preferable, preferably solves biosensor with depositing
The problem that the background current that the storage time increases and increases raises, so that the storage stability of biosensor is more preferably, it is ensured that
Biosensor test accuracy before the deadline.
Claims (10)
1. a biosensor, it is characterised in that composition includes insulated substrate, the electrode system being attached on insulated substrate, institute
The electrode system stated at least includes that a working electrode and one, to electrode, cover the reaction reagent layer on electrode system,
Described reaction reagent layer should at least cover a working electrode, and is positioned at the reaction above electrode system and reaction reagent layer
Room, described reative cell is made up of middle interlayer and cover layer, and reative cell has air-vent, wherein: described reaction reagent layer
Containing one or more in imidazoles or imidazole derivative in the reaction reagent used.
A kind of biosensor the most according to claim 1, it is characterised in that described imidazoles or imidazole derivative exist
Addition in reaction reagent is 1mM ~ 2M.
A kind of biosensor the most according to claim 1, it is characterised in that described imidazole derivative is selected from 1-first
Base imidazoles, 1-ethyl imidazol(e), 1-propyl imidazole, 2,4-methylimidazole, 2-methylimidazole, 2-ethyl imidazol(e), 4-methylimidazole,
DMIZ 1,2 dimethylimidazole, 2-ethyl-4-methylimidazole and 1-ethoxy-2-methylimidazole.
A kind of biosensor the most according to claim 1, it is characterised in that described reaction reagent includes that glucose is anti-
Answer reagent, lentochol reaction reagent, uric acid reaction reagent, ketoboidies reaction reagent, lactic acid reaction reagent and hemoglobin reaction examination
Agent.
A kind of biosensor the most according to claim 1, it is characterised in that the thickness of described middle interlayer is
0.025mm~0.5mm。
6. according to the preparation method of a kind of biosensor one of claim 1-5 Suo Shu, it is characterised in that including:
(1) in insulated base material layer formed electrode system, described electrode system at least include a working electrode and one right
Electrode, processing method has: by silk screen printing, specific electrode pattern is printed onto in insulated base material layer, then by heating or
Infrared or ultraviolet is dried；Or electrode material is attached in insulated base material layer by the way of sputtering, then passes through laser ablation
Mode process specific electrode pattern,
(2) reagent layer is fixed on electrode by the processing of a liquid, silk screen printing, inkjet printing or crack extrusion method,
(3) space layer of reative cell of fitting on electrode system and reagent layer and cover layer, complete, wherein: with band base material
Or the adhesive tape without base material is space layer material, it is bonded in after processing on electrode system and reagent layer；Or with glue or polymerization
Thing slurry is space layer material, by being screen printed onto on electrode system and reagent layer.
7. the reaction reagent of a biosensor, it is characterised in that derive containing imidazoles or imidazoles in described reaction reagent
One or more in thing.
The reaction reagent of a kind of biosensor the most according to claim 7, it is characterised in that described imidazoles or imidazoles
Analog derivative concentration interval in reaction reagent is 1mM ~ 2M.
The reaction reagent of a kind of biosensor the most according to claim 7, it is characterised in that described imidazoles derives
Thing selected from 1-Methylimidazole., 1-ethyl imidazol(e), 1-propyl imidazole, 2,4-methylimidazole, 2-methylimidazole, 2-ethyl imidazol(e),
4-methylimidazole, DMIZ 1,2 dimethylimidazole, 2-ethyl-4-methylimidazole and 1-ethoxy-2-methylimidazole.
The reaction reagent of a kind of biosensor the most according to claim 7, it is characterised in that described reaction reagent bag
Include glucose response reagent, lentochol reaction reagent, uric acid reaction reagent, ketoboidies reaction reagent, lactic acid reaction reagent and blood red
Albumino reaction reagent 2.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610436485.8A CN106124595A (en) | 2016-06-17 | 2016-06-17 | A kind of biosensor and preparation method thereof and reaction reagent |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610436485.8A CN106124595A (en) | 2016-06-17 | 2016-06-17 | A kind of biosensor and preparation method thereof and reaction reagent |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106124595A true CN106124595A (en) | 2016-11-16 |
Family
ID=57471159
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610436485.8A Pending CN106124595A (en) | 2016-06-17 | 2016-06-17 | A kind of biosensor and preparation method thereof and reaction reagent |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106124595A (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107064261A (en) * | 2017-05-01 | 2017-08-18 | 台州亿联健医疗科技有限公司 | A kind of biology sensor and detection method based on glucose dehydrogenase |
| CN107121478A (en) * | 2017-05-01 | 2017-09-01 | 台州亿联健医疗科技有限公司 | A kind of biology sensor that can detect that sample is not enough, method |
| CN107290417A (en) * | 2017-06-15 | 2017-10-24 | 浙江大学 | A kind of biology sensor for improving reagent crystal habit |
| CN107941880A (en) * | 2017-11-16 | 2018-04-20 | 江苏鱼跃医疗设备股份有限公司 | For improving the reaction reagent and glucose sensor comprising betanin derivative of glucose sensor storage stability |
| CN108760854A (en) * | 2018-05-07 | 2018-11-06 | 杭州电子科技大学 | A kind of preparation method of polynary no enzyme electrochemical glucose sensing material |
| TWI644101B (en) * | 2017-08-29 | 2018-12-11 | 瑞瑩生醫科技股份有限公司 | A test strip for electrochemical biosensor with high stability and manufacturing method thereof |
| CN114923965A (en) * | 2022-03-18 | 2022-08-19 | 杭州微策生物技术股份有限公司 | Integrated multi-index sensor and manufacturing method thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1462367A (en) * | 2001-05-15 | 2003-12-17 | 松下电器产业株式会社 | Biosensor |
| CN1875114A (en) * | 2003-10-29 | 2006-12-06 | 新加坡科技研究局 | Biosensor |
| CN101801985A (en) * | 2007-06-19 | 2010-08-11 | 霍夫曼-拉罗奇有限公司 | redox mediators |
| CN102027623A (en) * | 2008-05-15 | 2011-04-20 | 索尼公司 | Fuel cell, method for production of fuel cell, electronic device, enzyme-immobilized electrode, biosensor, bioreactor, energy conversion element, and enzymatic reaction-utilizing apparatus |
| CN102759552A (en) * | 2011-04-26 | 2012-10-31 | 爱科来株式会社 | Analysis device |
| CN103154717A (en) * | 2010-09-30 | 2013-06-12 | 松下电器产业株式会社 | Reagent composition, sensor, sensor system, and sensor manufacture process |
-
2016
- 2016-06-17 CN CN201610436485.8A patent/CN106124595A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1462367A (en) * | 2001-05-15 | 2003-12-17 | 松下电器产业株式会社 | Biosensor |
| CN1875114A (en) * | 2003-10-29 | 2006-12-06 | 新加坡科技研究局 | Biosensor |
| CN101801985A (en) * | 2007-06-19 | 2010-08-11 | 霍夫曼-拉罗奇有限公司 | redox mediators |
| CN102027623A (en) * | 2008-05-15 | 2011-04-20 | 索尼公司 | Fuel cell, method for production of fuel cell, electronic device, enzyme-immobilized electrode, biosensor, bioreactor, energy conversion element, and enzymatic reaction-utilizing apparatus |
| CN103154717A (en) * | 2010-09-30 | 2013-06-12 | 松下电器产业株式会社 | Reagent composition, sensor, sensor system, and sensor manufacture process |
| CN102759552A (en) * | 2011-04-26 | 2012-10-31 | 爱科来株式会社 | Analysis device |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107064261A (en) * | 2017-05-01 | 2017-08-18 | 台州亿联健医疗科技有限公司 | A kind of biology sensor and detection method based on glucose dehydrogenase |
| CN107121478A (en) * | 2017-05-01 | 2017-09-01 | 台州亿联健医疗科技有限公司 | A kind of biology sensor that can detect that sample is not enough, method |
| CN107290417A (en) * | 2017-06-15 | 2017-10-24 | 浙江大学 | A kind of biology sensor for improving reagent crystal habit |
| TWI644101B (en) * | 2017-08-29 | 2018-12-11 | 瑞瑩生醫科技股份有限公司 | A test strip for electrochemical biosensor with high stability and manufacturing method thereof |
| CN107941880A (en) * | 2017-11-16 | 2018-04-20 | 江苏鱼跃医疗设备股份有限公司 | For improving the reaction reagent and glucose sensor comprising betanin derivative of glucose sensor storage stability |
| CN107941880B (en) * | 2017-11-16 | 2020-07-31 | 江苏鱼跃医疗设备股份有限公司 | Reaction reagent for improving storage stability of glucose sensor comprising betaine derivative, and glucose sensor |
| CN108760854A (en) * | 2018-05-07 | 2018-11-06 | 杭州电子科技大学 | A kind of preparation method of polynary no enzyme electrochemical glucose sensing material |
| CN114923965A (en) * | 2022-03-18 | 2022-08-19 | 杭州微策生物技术股份有限公司 | Integrated multi-index sensor and manufacturing method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106124595A (en) | A kind of biosensor and preparation method thereof and reaction reagent | |
| Jia et al. | Engineering the bioelectrochemical interface using functional nanomaterials and microchip technique toward sensitive and portable electrochemical biosensors | |
| Sakata | Biologically coupled gate field-effect transistors meet in vitro diagnostics | |
| Poletti et al. | Continuous capillary-flow sensing of glucose and lactate in sweat with an electrochemical sensor based on functionalized graphene oxide | |
| Kim et al. | A highly permselective electrochemical glucose sensor using red blood cell membrane | |
| US20150122646A1 (en) | Mediator-less Electrochemical Glucose Sensing Procedure Employing the Leach-proof Covalent Binding of an Enzyme(s) to Electrodes and Products Thereof | |
| Cunha-Silva et al. | Dual range lactate oxidase-based screen printed amperometric biosensor for analysis of lactate in diversified samples | |
| Bhardwaj | A review on immobilization techniques of biosensors | |
| CN107064261A (en) | A kind of biology sensor and detection method based on glucose dehydrogenase | |
| Çolak et al. | Glucose biosensor based on the immobilization of glucose oxidase on electrochemically synthesized polypyrrole-poly (vinyl sulphonate) composite film by cross-linking with glutaraldehyde | |
| CN104931560A (en) | Biosensor and preparation method and application thereof | |
| Kekec et al. | A novel conducting polymer based platform for ethanol sensing | |
| Li et al. | Recent advances in enzyme inhibition based-electrochemical biosensors for pharmaceutical and environmental analysis | |
| CN107121478A (en) | A kind of biology sensor that can detect that sample is not enough, method | |
| CN112014448A (en) | Biosensor, method for manufacturing the same, and polymer film layer for biosensor | |
| AU2019283368B2 (en) | Crosslinker comprising genipin for use in preparation of sensing film or diffusion control film of electrochemical sensor | |
| Feng et al. | A disposable cholesterol enzyme biosensor based on ferrocene-capped gold nanoparticle modified screen-printed carbon electrode | |
| Zhang et al. | Enzyme powered self-assembly of hydrogel biosensor for colorimetric detection of metabolites | |
| Wayu et al. | Layered Xerogel Films Incorporating Monolayer‐Protected Cluster Networks on Platinum‐Black‐Modified Electrodes for Enhanced Sensitivity in First‐Generation Uric Acid Biosensing | |
| Kurbanoglu et al. | Nanobiodevices for electrochemical biosensing of pharmaceuticals | |
| KR20200003008A (en) | Biosensors Produced from Enzymes with Reduced Solubility and Methods for Producing and Using Them | |
| CA3055183C (en) | Nanobead containing biosensors and methods of production and use thereof | |
| Kucherenko et al. | A novel adenosine triphosphate (ATP) biosensor based on electrospun polymer nanofibers with entrapped hexokinase and glucose oxidase | |
| Chen et al. | A high selective disposable biosensor based on screen-printed technique with two working electrodes for eliminating interference signals | |
| CN208795691U (en) | A kind of beta-hydroxybutyric acid electrochemical sensor |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20161116 |
|
| RJ01 | Rejection of invention patent application after publication |