CN104792832A - Bilirubin electrochemical transducer and preparation method thereof - Google Patents
Bilirubin electrochemical transducer and preparation method thereof Download PDFInfo
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- CN104792832A CN104792832A CN201510180693.1A CN201510180693A CN104792832A CN 104792832 A CN104792832 A CN 104792832A CN 201510180693 A CN201510180693 A CN 201510180693A CN 104792832 A CN104792832 A CN 104792832A
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Abstract
The invention relates to the technical field of electrochemical detection, in particular to a bilirubin electrochemical transducer and a preparation method thereof. The bilirubin electrochemical transducer comprises an electrode, a bilirubin oxidase layer and an anti-interference enzyme layer, wherein the bilirubin oxidase layer is arranged between the electrode and the anti-interference enzyme layer, and comprises bilirubin oxidase and an oxidation-state electronic mediator; the anti-interference enzyme layer comprises ascorbic acid oxidase. The bilirubin electrochemical transducer has the advantages of being high in sensitivity, wide in linearity range, low in sample demand, high in anti-interference capacity, and short in detection time.
Description
Technical field
The present invention relates to technical field of electrochemical detection, particularly a kind of cholerythrin electrochemical sensor and preparation method thereof.
Background technology
Cholerythrin is the one of BILE PIGMENTS, and it is the primary pigments in human bile, in orange-yellow.It is the main metabolites of iron porphyrin compound in body, toxic, can cause irreversible infringement, but also have antioxidant function, can suppress the oxidation of linoleic acid and phosphatide to brain and nervous system.Cholerythrin is the important evidence judging jaundice clinically, is also the important indicator of liver function.Such as: when liver cell generation pathology or because of (be mostly suffer from the liver diseases such as acute icteric hepatitis, acute yellow hepatonecrosis, chronic active hepatitis, cirrhosis cause) during swelling of liver cell, the bile duct pressurized in liver can be caused, excretion of bile is obstructed, thus the phenomenon that blood mesobilirubin can be caused higher, and there is parenchymal jaundice (show as bilirubin direct and indirect bilirubin all raises); When the biliary system generation tumour outside liver or occur calculus, and during by biliary obstruction, bile can not be drained smoothly, and cholerythrin can be caused higher, and obstructive jaundice occurs.
Current cholerythrin detects and mainly contains diazo reagent method, chemical oxidization method, bilirubin oxidase end-point method:
The application of diazo reagent method the earliest, its Cleaning Principle is: in serum combined with bilirubin can directly and diazo reagent react and produce azobilirubin, will with caffeine when unconjugated bilirubin measures, Sodium Benzoate is that accelerator destruction cholerythrin hydrogen bond reacts with diazo reagent again, ascorbic acid destroys residue diazo reagent, adding alkaline sodium tartrate makes maximum absorbance forward 598nm to by 530nm, non-bilirubinic xanthein and other red absorbance produced with brown pigmentation are down to and can be ignored, sensitivity and specificity are raised, the green finally formed is mixed by the alkaline azobilirubin of blue look and the xanthein that formed between caffeine and sulfanilic acid.But the method range of linearity is narrower, difficult quality controls, and result difference is large;
Chemical oxidization method developed recently is rapid, and improvement chemical oxidization method adopts inorganic oxidizer oxidation cholerythrin, causes the decline of 450nm wavelength absorbance.But the method is easily disturbed, often there is negative value in result;
1987; Kosaka and Doumas reports the enzymatic assays of serum direct bilirubin respectively; experiment is using ditaurobilirubin as bilirubin direct standard; reactant liquor pH is controlled at acid range; with this understanding, the glucuronic acid combined with bilirubin in serum and most of protein combination cholerythrin all oxidized, cholerythrin under the effect of bilirubin oxidase with oxygen reaction; generate dehydrobilirubin, dehydrobilirubin with water effect.Cholerythrin maximum absorption band is near 450nm, and under the effect of bilirubin oxidase, total bilirubin is directly proportional to cholerythrin in the decline of 450nm absorbance.The Bilirubin Oxidase method range of linearity is wide, but bilirubin oxidase end-point method computing formula is complicated, and the reaction time is long, is subject to the interference of serum background.
Above-mentioned three kinds of cholerythrin detection methods are all photochemical methods, all there is the problem that antijamming capability is poor.Therefore, a kind of the cholerythrin detection method and the reagent thereof that improve antijamming capability is badly in need of.
Summary of the invention
In view of this, the invention provides a kind of cholerythrin electrochemical sensor and preparation method thereof.This cholerythrin electrochemical sensor have highly sensitive, the range of linearity is wide, need the advantages such as sample amount is few, antijamming capability is strong, detection time is short.
In order to realize foregoing invention object, the invention provides following technical scheme:
The invention provides a kind of cholerythrin electrochemical sensor, comprise electrode, bilirubin oxidase layer and anti-interference enzyme layer, bilirubin oxidase layer is arranged between electrode and anti-interference enzyme layer;
Bilirubin oxidase layer comprises bilirubin oxidase, oxidation state electron mediator;
Anti-interference enzyme layer comprises ascorbic acid oxidase.
Cleaning Principle of the present invention is: use electrochemical test method, by bilirubin oxidase (oxidation state), the cholerythrin in sample is oxidized to dehydrobilirubin, bilirubin oxidase (oxidation state) is transformed into bilirubin oxidase (reduction-state), electron mediator (oxidation state) is reduced into electron mediator (reduction-state) by bilirubin oxidase (reduction-state), when adding positive voltage, electron mediator (reduction-state) is oxidized to electron mediator (oxidation state), generation current signal simultaneously, current signal strength is directly proportional to bilirubin concentration (linear), bilirubin concentration can be obtained by sensed current signal.Reaction equation is as follows:
Cholerythrin+bilirubin oxidase (oxidation state) → dehydrobilirubin+bilirubin oxidase (reduction-state)
Electron mediator (oxidation state)+bilirubin oxidase (reduction-state) → electron mediator (reduction-state)+bilirubin oxidase (oxidation state)
Electron mediator (reduction-state)+voltage → electron mediator (oxidation state)+electric current
Research in the past finds that Bilirubin Oxidase method exists the situation being subject to the interference of serum background, but does not learn concrete disturbing factor.The present invention studies discovery, and when utilizing Bilirubin Oxidase method to detect the content of sample mesobilirubin, why this method is subject to the interference of serum background, and reason is in sample containing interfering material ascorbic acid.The present invention is by be printed on anti-interference enzyme layer on electrode and to be covered in the upper strata of bilirubin oxidase layer, thus make before sample and bilirubin oxidase (oxidation state) react, sample can first through anti-interference enzyme layer, interfering material ascorbic acid in sample is consumed by reacting with the ascorbic acid oxidase in anti-interference enzyme layer, thus promotes the accuracy detected.
In the present invention, bilirubin oxidase is the bilirubin oxidase being in oxidation state, has oxidisability.
In embodiments more provided by the invention, the enzyme activity of bilirubin oxidase is 10 ~ 100U/mg.
As preferably, the enzyme activity of bilirubin oxidase is 14U/mg.
In the present invention, electron mediator is the electron mediator being in oxidation state, has oxidisability.
In the present invention, ascorbic acid oxidase is the ascorbic acid oxidase being in oxidation state.
In embodiments more provided by the invention, the enzyme activity of ascorbic acid oxidase is 10 ~ 200U/mg.
As preferably, the enzyme activity of ascorbic acid oxidase is 100U/mg.
In embodiments more provided by the invention, bilirubin oxidase layer also comprises spreading agent and macromolecule film former, and anti-interference enzyme layer also comprises surfactant and macromolecule film former.
As preferably, with parts by weight, 0.1 ~ 5 part, bilirubin oxidase layer mesobilirubin oxidase, oxidation state electron mediator 1 ~ 15 part, spreading agent 0.1 ~ 10 part, macromolecule film former 10 ~ 50 parts;
Ascorbic acid oxidase 1 ~ 5 part in anti-interference enzyme layer, 1 ~ 10 part, surfactant, macromolecule film former 10 ~ 50 parts.
Preferably, with parts by weight, 3 parts, bilirubin oxidase layer mesobilirubin oxidase, oxidation state electron mediator 10 parts, spreading agent 5 parts, macromolecule film former 20 parts;
Ascorbic acid oxidase 2 parts in anti-interference enzyme layer, 5 parts, surfactant, macromolecule film former 20 parts.
Electron mediator is also known as mediator (electron transfer mediator, electro transfer mediator, electron mediator etc.), to refer to the electronics that produces in enzyme reaction process from enzyme reaction central transference to electrode surface, make electrode produce point electronic conductor of corresponding curent change.In embodiments more provided by the invention, electron mediator is a kind of or both the above potpourris in tri-chlorination six ammino ruthenium, cupric chloride, copper sulphate, Prussian blue, adjacent luxuriant and rich with fragrance quinoline and derivant thereof, methylene blue.But the kind of electron mediator is not defined in this, the electron mediator of those skilled in the art's accreditation is all within protection scope of the present invention, and the present invention is in this no limit.
In embodiments more provided by the invention, spreading agent is a kind of or both the above potpourris in EFKA 4320, Dispers 655, BYK163.But the kind of spreading agent is not defined in this, the spreading agent of those skilled in the art's accreditation is all within protection scope of the present invention, and the present invention is in this no limit.
In embodiments more provided by the invention, surfactant is a kind of or both the above potpourris in tween, Qu Latong, lauryl sodium sulfate, polyglycol.But the kind of surfactant is not defined in this, the surfactant of those skilled in the art's accreditation is all within protection scope of the present invention, and the present invention is in this no limit.
In embodiments more provided by the invention, macromolecule film former is a kind of or both the above potpourris in carboxymethyl cellulose, hydroxyethyl cellulose, shitosan, gelatin.But the kind of macromolecule film former is not defined in this, the macromolecule film former of those skilled in the art's accreditation is all within protection scope of the present invention, and the present invention is in this no limit.
In embodiments more provided by the invention, electrode is carbon electrode.But the kind of electrode is not defined in this, the electrode of those skilled in the art's accreditation is all within protection scope of the present invention, and the present invention is in this no limit.
Present invention also offers a kind of preparation method of cholerythrin electrochemical sensor, comprise the steps:
Bilirubin oxidase, oxidation state electron mediator, spreading agent, solvent and macromolecule film former are mixed, obtains bilirubin oxidase system;
Ascorbic acid oxidase, surfactant, solvent and macromolecule film former are mixed, obtains anti-interference enzyme system;
Bilirubin oxidase system, anti-interference enzyme system are printed on electrode successively, obtain cholerythrin electrochemical sensor.
In preparation method of the present invention, bilirubin oxidase is the bilirubin oxidase being in oxidation state, has oxidisability.
In embodiments more provided by the invention, the enzyme activity of bilirubin oxidase is 10 ~ 100U/mg.
As preferably, the enzyme activity of bilirubin oxidase is 14U/mg.
In preparation method of the present invention, electron mediator is the electron mediator being in oxidation state, has oxidisability.
In preparation method of the present invention, ascorbic acid oxidase is the ascorbic acid oxidase being in oxidation state.
In embodiments more provided by the invention, the enzyme activity of ascorbic acid oxidase is 10 ~ 200U/mg.
As preferably, the enzyme activity of ascorbic acid oxidase is 100U/mg.
In the preparation method of cholerythrin electrochemical sensor, with parts by weight, 0.1 ~ 5 part, bilirubin oxidase layer mesobilirubin oxidase, oxidation state electron mediator 1 ~ 15 part, spreading agent 0.1 ~ 10 part, solvent 950 ~ 990 parts, macromolecule film former 10 ~ 50 parts;
Ascorbic acid oxidase 1 ~ 5 part in anti-interference enzyme layer, 1 ~ 10 part, surfactant, solvent 950 ~ 990 parts, macromolecule film former 10 ~ 50 parts.
Preferably, with parts by weight, 3 parts, bilirubin oxidase layer mesobilirubin oxidase, oxidation state electron mediator 10 parts, spreading agent 5 parts, solvent 970 parts, macromolecule film former 20 parts;
Ascorbic acid oxidase 2 parts in anti-interference enzyme layer, 5 parts, surfactant, solvent 980 parts, macromolecule film former 20 parts.
In embodiments more provided by the invention, electron mediator is a kind of or both the above potpourris in tri-chlorination six ammino ruthenium, cupric chloride, copper sulphate, Prussian blue, adjacent luxuriant and rich with fragrance quinoline and derivant thereof, methylene blue.
In embodiments more provided by the invention, spreading agent is a kind of or both the above potpourris in EFKA 4320, Dispers 655, BYK163.
In embodiments more provided by the invention, surfactant is a kind of or both the above potpourris in tween, Qu Latong, lauryl sodium sulfate, polyglycol.
In embodiments more provided by the invention, macromolecule film former is a kind of or both the above potpourris in carboxymethyl cellulose, hydroxyethyl cellulose, shitosan, gelatin.
In embodiments more provided by the invention, electrode is carbon electrode.But the kind of electrode is not defined in this, the electrode of those skilled in the art's accreditation is all within protection scope of the present invention, and the present invention is in this no limit.
As preferably, print for serigraphy.
In embodiments more provided by the invention, solvent is a kind of or both the above potpourris in ethanol, isopropyl alcohol, ethyl acetate, water.
Present invention also offers the bilirubinic detection method of a kind of non-diagnostic object, comprise the steps:
After sample and ascorbic acid oxidase being reacted, adopt Bilirubin Oxidase method sensed current signal, according to the concentration of current signal determination sample mesobilirubin.
The invention provides a kind of cholerythrin electrochemical sensor and preparation method thereof.This cholerythrin electrochemical sensor comprises electrode, bilirubin oxidase layer and anti-interference enzyme layer, and bilirubin oxidase layer is arranged between electrode and anti-interference enzyme layer; Bilirubin oxidase layer comprises bilirubin oxidase, oxidation state electron mediator; Anti-interference enzyme layer comprises ascorbic acid oxidase.The present invention at least comprises one of following advantage:
The present invention is by be printed on anti-interference enzyme layer on electrode and to be covered in the upper strata of bilirubin oxidase layer, thus make before sample and bilirubin oxidase (oxidation state) react, sample can first through anti-interference enzyme layer, interfering material ascorbic acid in sample is consumed by reacting with the ascorbic acid oxidase in anti-interference enzyme layer, thus promoting the accuracy detected, antijamming capability is strong;
This cholerythrin electrochemical sensor have simultaneously highly sensitive, the range of linearity is wide, need the advantages such as sample amount is few, detection time is short.
Accompanying drawing explanation
Fig. 1 shows the corresponding relation of cholerythrin and current value; Wherein, series 1 shows the current value test curve of cholerythrin electrochemical sensor provided by the invention under various biliary lycopene concentration.
Embodiment
The invention discloses a kind of cholerythrin electrochemical sensor and preparation method thereof, those skilled in the art can use for reference present disclosure, and suitable improving technique parameter realizes.Special needs to be pointed out is, all similar replacements and change apparent to those skilled in the art, they are all deemed to be included in the present invention.Method of the present invention and application are described by preferred embodiment, related personnel obviously can not depart from content of the present invention, spirit and scope methods and applications as herein described are changed or suitably change with combination, realize and apply the technology of the present invention.
In cholerythrin electrochemical sensor provided by the invention and preparation method thereof, agents useful for same, material all can be buied by market.
Below in conjunction with embodiment, set forth the present invention further:
The preparation of embodiment 1 cholerythrin electrochemical sensor
Use screen printing technique to be printed on PET by conductive carbon paste and be made into carbon electrode.
Take the copper sulphate of 1g, the bilirubin oxidase of 0.01g, BYK163,2g carboxymethyl cellulose of 0.5g, 97g pure water, use homo-mixer stirs while adding, rotating speed controls at 600rpm, time controling was at 15 minutes, Deng enzyme liquid mix after stirring machine rotating speed is controlled to stir 3 hours at 1200rpm again, obtain bilirubin oxidase system enzyme liquid; Use screen printing technique to be printed on carbon electrode obtained enzyme liquid, by oven for drying film forming, obtain cholerythrin electrochemical sensor semi-manufacture.
Take 0.2g ascorbic acid oxidase, 0.5g Tween-60,2g carboxymethyl cellulose, 98g pure water, use homo-mixer stirs while adding, rotating speed controls at 600rpm, time controling was at 15 minutes, again stirring machine rotating speed is controlled to stir 3 hours at 1200rpm after mixing Deng enzyme liquid, obtain anti-interference enzyme system enzyme liquid; Use screen printing technique to be printed on cholerythrin electrochemical sensor semi-manufacture obtained enzyme liquid, by oven for drying film forming, cut, roller shear, packing, obtain cholerythrin electrochemical sensor.
Use electrochemical workstation testing sensor time current curve under variable concentrations cholerythrin (1mg/dl ~ 40mg/dl), its size of current and bilirubin concentration linear.Test findings is in table 1, Fig. 1.
The current value test chart of table 1 sensor under various biliary lycopene concentration
| Bilirubin concentration (mg/dl) | 1 | 4 | 9 | 28 | 40 |
| Current value (nA) | 137.0 | 165.0 | 230.0 | 396.0 | 560.0 |
The preparation of embodiment 2 cholerythrin electrochemical sensor
Use screen printing technique to be printed on PET by conductive carbon paste and be made into carbon electrode.
Take the cupric chloride of 1.5g, the bilirubin oxidase of 15mg, the TEGO Dispers 655 of 0.5g, 2g hydroxyethyl cellulose, 66g pure water, 30g isopropyl alcohol, use homo-mixer stirs while adding, rotating speed controls at 600rpm, time controling was at 15 minutes, Deng enzyme liquid mix after stirring machine rotating speed is controlled to stir 3 hours at 1200rpm again, obtain bilirubin oxidase system enzyme liquid; Use screen printing technique to be printed on carbon electrode obtained enzyme liquid, by oven for drying film forming, obtain cholerythrin electrochemical sensor semi-manufacture.
Take 0.1g ascorbic acid oxidase, 0.5g song draws logical, 5g hydroxyethyl cellulose, 95g pure water, use homo-mixer stirs while adding, rotating speed controls at 600rpm, time controling was at 15 minutes, again stirring machine rotating speed is controlled to stir 3 hours at 1200rpm after mixing Deng enzyme liquid, obtain anti-interference enzyme system enzyme liquid; Use screen printing technique to be printed on cholerythrin electrochemical sensor semi-manufacture obtained enzyme liquid, by oven for drying film forming, cut, roller shear, packing, obtain cholerythrin electrochemical sensor.
Use electrochemical workstation testing sensor time current curve under variable concentrations cholerythrin (1mg/dl ~ 40mg/dl), its size of current and bilirubin concentration linear.
The preparation of embodiment 3 cholerythrin electrochemical sensor
Use screen printing technique to be printed on PET by conductive carbon paste and be made into carbon electrode.
Take the methylene blue of 1.5g, the bilirubin oxidase of 15mg, EFKA4320,2g shitosan of 1g, 75g pure water, 20g ethanol, use homo-mixer stirs while adding, rotating speed controls at 600rpm, time controling was at 15 minutes, Deng enzyme liquid mix after stirring machine rotating speed is controlled to stir 3 hours at 1200rpm again, obtain bilirubin oxidase system enzyme liquid; Use screen printing technique to be printed on carbon electrode obtained enzyme liquid, by oven for drying film forming, obtain cholerythrin electrochemical sensor semi-manufacture.
Take 0.3g ascorbic acid oxidase, 1g lauryl sodium sulfate, 1g shitosan, 98g pure water, use homo-mixer stirs while adding, rotating speed controls at 600rpm, time controling was at 15 minutes, again stirring machine rotating speed is controlled to stir 3 hours at 1200rpm after mixing Deng enzyme liquid, obtain anti-interference enzyme system enzyme liquid; Use screen printing technique to be printed on cholerythrin electrochemical sensor semi-manufacture obtained enzyme liquid, by oven for drying film forming, cut, roller shear, packing, obtain cholerythrin electrochemical sensor.
Use electrochemical workstation testing sensor time current curve under variable concentrations cholerythrin (1mg/dl ~ 40mg/dl), its size of current and bilirubin concentration linear.
The preparation of embodiment 4 cholerythrin electrochemical sensor
Use screen printing technique to be printed on PET by conductive carbon paste and be made into carbon electrode.
Take the tri-chlorination six ammino ruthenium of 1g, the bilirubin oxidase of 0.3g, TEGO Dispers655,2g gelatin of 0.5g, 67g pure water, 30g ethyl acetate, use homo-mixer stirs while adding, rotating speed controls at 600rpm, time controling was at 15 minutes, Deng enzyme liquid mix after stirring machine rotating speed is controlled to stir 3 hours at 1200rpm again, obtain bilirubin oxidase system enzyme liquid; Use screen printing technique to be printed on carbon electrode obtained enzyme liquid, by oven for drying film forming, obtain cholerythrin electrochemical sensor semi-manufacture.
Take 0.5g ascorbic acid oxidase, 0.1g polyglycol, 3g gelatin, 67g pure water, 30g ethyl acetate, use homo-mixer stirs while adding, rotating speed controls at 600rpm, time controling was at 15 minutes, again stirring machine rotating speed is controlled to stir 3 hours at 1200rpm after mixing Deng enzyme liquid, obtain anti-interference enzyme system enzyme liquid; Use screen printing technique to be printed on cholerythrin electrochemical sensor semi-manufacture obtained enzyme liquid, by oven for drying film forming, cut, roller shear, packing, obtain cholerythrin electrochemical sensor.
Use electrochemical workstation testing sensor time current curve under variable concentrations cholerythrin (1mg/dl ~ 40mg/dl), its size of current and bilirubin concentration linear.
The preparation of embodiment 5 cholerythrin electrochemical sensor
Use screen printing technique to be printed on PET by conductive carbon paste and be made into carbon electrode.
Take Prussian blue, the bilirubin oxidase of 10mg, the BYK 163 of 1g, 1g carboxymethyl cellulose, the 95g pure water of 1.5g, use homo-mixer stirs while adding, rotating speed controls at 600rpm, time controling was at 15 minutes, Deng enzyme liquid mix after stirring machine rotating speed is controlled to stir 3 hours at 1200rpm again, obtain bilirubin oxidase system enzyme liquid; Use screen printing technique to be printed on carbon electrode obtained enzyme liquid, by oven for drying film forming, obtain cholerythrin electrochemical sensor semi-manufacture.
Take 0.4g ascorbic acid oxidase, 0.8g Tween-80,4g carboxymethyl cellulose, 70g pure water, 25g ethanol, use homo-mixer stirs while adding, rotating speed controls at 600rpm, time controling was at 15 minutes, again stirring machine rotating speed is controlled to stir 3 hours at 1200rpm after mixing Deng enzyme liquid, obtain anti-interference enzyme system enzyme liquid; Use screen printing technique to be printed on cholerythrin electrochemical sensor semi-manufacture obtained enzyme liquid, by oven for drying film forming, cut, roller shear, packing, obtain cholerythrin electrochemical sensor.
Use electrochemical workstation testing sensor time current curve under variable concentrations cholerythrin (1mg/dl ~ 40mg/dl), its size of current and bilirubin concentration linear.
The preparation of embodiment 6 cholerythrin electrochemical sensor
Use screen printing technique to be printed on PET by conductive carbon paste and be made into carbon electrode.
Take neighbour luxuriant and rich with fragrance quinoline, the bilirubin oxidase of 0.5g, EFKA4320,5g hydroxyethyl cellulose of 0.01g, the 95g pure water of 0.1g, use homo-mixer stirs while adding, rotating speed controls at 600rpm, time controling was at 15 minutes, Deng enzyme liquid mix after stirring machine rotating speed is controlled to stir 3 hours at 1200rpm again, obtain bilirubin oxidase system enzyme liquid; Use screen printing technique to be printed on carbon electrode obtained enzyme liquid, by oven for drying film forming, obtain cholerythrin electrochemical sensor semi-manufacture.
Take 0.2g ascorbic acid oxidase, 0.6g song draws logical, 4.5g hydroxyethyl cellulose, 95g pure water, use homo-mixer stirs while adding, rotating speed controls at 600rpm, time controling was at 15 minutes, again stirring machine rotating speed is controlled to stir 3 hours at 1200rpm after mixing Deng enzyme liquid, obtain anti-interference enzyme system enzyme liquid; Use screen printing technique to be printed on cholerythrin electrochemical sensor semi-manufacture obtained enzyme liquid, by oven for drying film forming, cut, roller shear, packing, obtain cholerythrin electrochemical sensor.
Use electrochemical workstation testing sensor time current curve under variable concentrations cholerythrin (1mg/dl ~ 40mg/dl), its size of current and bilirubin concentration linear.
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the premise without departing from the principles of the invention; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (10)
1. a cholerythrin electrochemical sensor, is characterized in that, comprises electrode, bilirubin oxidase layer and anti-interference enzyme layer, and described bilirubin oxidase layer is arranged between described electrode and described anti-interference enzyme layer;
Described bilirubin oxidase layer comprises bilirubin oxidase, oxidation state electron mediator;
Described anti-interference enzyme layer comprises ascorbic acid oxidase.
2. cholerythrin electrochemical sensor according to claim 1, is characterized in that, described bilirubin oxidase layer also comprises spreading agent and macromolecule film former, and described anti-interference enzyme layer also comprises surfactant and macromolecule film former.
3. cholerythrin electrochemical sensor according to claim 2, is characterized in that, with parts by weight, described 0.1 ~ 5 part, bilirubin oxidase layer mesobilirubin oxidase, oxidation state electron mediator 1 ~ 15 part, spreading agent 0.1 ~ 10 part, macromolecule film former 10 ~ 50 parts;
Ascorbic acid oxidase 1 ~ 5 part in described anti-interference enzyme layer, 1 ~ 10 part, surfactant, macromolecule film former 10 ~ 50 parts.
4. cholerythrin electrochemical sensor according to claim 1, is characterized in that, electron mediator is a kind of or both the above potpourris in tri-chlorination six ammino ruthenium, cupric chloride, copper sulphate, Prussian blue, adjacent luxuriant and rich with fragrance quinoline and derivant thereof, methylene blue.
5. cholerythrin electrochemical sensor according to claim 2, is characterized in that, described spreading agent is a kind of or both the above potpourris in EFKA 4320, Dispers 655, BYK 163.
6. cholerythrin electrochemical sensor according to claim 2, is characterized in that, described surfactant is a kind of or both the above potpourris in tween, Qu Latong, lauryl sodium sulfate, polyglycol.
7. cholerythrin electrochemical sensor according to claim 2, is characterized in that, described macromolecule film former is a kind of or both the above potpourris in carboxymethyl cellulose, hydroxyethyl cellulose, shitosan, gelatin.
8. a preparation method for cholerythrin electrochemical sensor, is characterized in that, comprises the steps:
Bilirubin oxidase, oxidation state electron mediator, spreading agent, solvent and macromolecule film former are mixed, obtains bilirubin oxidase system;
Ascorbic acid oxidase, surfactant, solvent and macromolecule film former are mixed, obtains anti-interference enzyme system;
Described bilirubin oxidase system, described anti-interference enzyme system are printed on electrode successively, obtain cholerythrin electrochemical sensor.
9. preparation method according to claim 8, is characterized in that, described printing is serigraphy.
10. the bilirubinic detection method of non-diagnostic object, is characterized in that, comprise the steps:
After sample and ascorbic acid oxidase being reacted, adopt Bilirubin Oxidase method sensed current signal, according to the concentration of described current signal determination sample mesobilirubin.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109580927A (en) * | 2019-01-04 | 2019-04-05 | 杭州联晟生物科技有限公司 | A kind of test card of liver function and preparation method thereof |
| CN111228627A (en) * | 2020-02-11 | 2020-06-05 | 中山大学附属第三医院 | Device and system for detecting intestinal fistula, bile fistula and pancreatic juice fistula |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1292086A (en) * | 1998-01-23 | 2001-04-18 | 艾－斯塔特公司 | Microfabricated aperture-based sensor |
| CN103038636A (en) * | 2010-06-07 | 2013-04-10 | 拜尔健康护理有限责任公司 | Underfill management system for a biosensor |
| 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 |
-
2015
- 2015-04-16 CN CN201510180693.1A patent/CN104792832A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1292086A (en) * | 1998-01-23 | 2001-04-18 | 艾－斯塔特公司 | Microfabricated aperture-based sensor |
| CN103038636A (en) * | 2010-06-07 | 2013-04-10 | 拜尔健康护理有限责任公司 | Underfill management system for a biosensor |
| 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 |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109580927A (en) * | 2019-01-04 | 2019-04-05 | 杭州联晟生物科技有限公司 | A kind of test card of liver function and preparation method thereof |
| CN111228627A (en) * | 2020-02-11 | 2020-06-05 | 中山大学附属第三医院 | Device and system for detecting intestinal fistula, bile fistula and pancreatic juice fistula |
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