CN110133079A - Glutamic-oxalacetic transaminease Electrochemical Detection composition, its application, electrochemical sensor and detection method - Google Patents
Glutamic-oxalacetic transaminease Electrochemical Detection composition, its application, electrochemical sensor and detection method Download PDFInfo
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- CN110133079A CN110133079A CN201910337020.0A CN201910337020A CN110133079A CN 110133079 A CN110133079 A CN 110133079A CN 201910337020 A CN201910337020 A CN 201910337020A CN 110133079 A CN110133079 A CN 110133079A
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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
- G01N27/3277—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction being a redox reaction, e.g. detection by cyclic voltammetry
Abstract
The invention discloses a kind of glutamic-oxalacetic transaminease Electrochemical Detection compositions, including first chamber and second chamber, the first chamber includes water, the first buffer, the first electrolyte, first surface activating agent, the first electron mediator and pyruvate oxidase, and the second chamber includes water, the second buffer, the second electrolyte, second surface activating agent, the second electron mediator, ASPARTIC ACID, l-Alanine, oxaloacetic decarboxylase and pyruvate oxidase.The invention also discloses a kind of applications of glutamic-oxalacetic transaminease Electrochemical Detection composition.The invention also discloses a kind of glutamic-oxalacetic transaminease electrochemical sensor and a kind of detection methods of glutamic-oxalacetic transaminease.
Description
Technical field
The present invention relates to field of biological detection, more particularly to a kind of glutamic-oxalacetic transaminease Electrochemical Detection composition, it is answered
With, electrochemical sensor and detection method.
Background technique
Glutamic-oxalacetic transaminease is the important enzyme of one kind of human metabolism.Glutamic-oxalacetic transaminease concentration in human body is quantified
Detection, it will be appreciated that the current hepatic disorder degree of human body and judge therapeutic effect, healthy population can also be carried out routinely
Liver function grade.
The general concentration by the detection characterization human body glutamic-oxalacetic transaminease of glutamic-oxalacetic transaminease in blood.Electrochemical sensor is
For detecting a kind of method of components in blood concentration.It can be by acquiring finger tip using electrochemical sensor detected components concentration
Blood is tested, and needs blood volume fewer, about 10 μ l or so, easy to operate, can carry out quantitative detection.Due to blood sampling volume
Few, requirement of the electrochemical sensor for precision is very high, and sensor can also compare the interference of non-target components in detection process
It is sensitive.And can have endogenic pyruvic acid in normal finger tip blood, the presence of pyruvic acid can make the detection of glutamic-oxalacetic transaminease
At interference, cause testing result inaccurate.
Summary of the invention
Based on this, it is necessary to which the interference of endogenous pyruvic acid can be excluded by providing one kind, and the millet straw for improving accuracy of detection turns
Adnosine deaminase Electrochemical Detection composition, its application, electrochemical sensor and detection method.
A kind of glutamic-oxalacetic transaminease Electrochemical Detection composition, including first chamber and second chamber, described first group
Closing object includes water, the first buffer, the first electrolyte, first surface activating agent, the first electron mediator and pyruvate oxidation
Enzyme, the second chamber include water, the second buffer, the second electrolyte, second surface activating agent, the second electron mediator,
ASPARTIC ACID, l-Alanine, oxaloacetic decarboxylase and pyruvate oxidase.
The concentration of the pyruvate oxidase in the first chamber is 0.1mg/mL in one of the embodiments,
~10mg/mL.
The concentration of the pyruvate oxidase in the second chamber is 0.1mg/mL in one of the embodiments,
~10mg/mL.
The concentration of the ASPARTIC ACID in the second chamber is 0.1mg/mL in one of the embodiments,
~10mg/mL, the concentration of the l-Alanine are 0.1mg/mL~10mg/mL.
The concentration of the oxaloacetic decarboxylase is 0.1mg/mL~10mg/mL in one of the embodiments,.
The second chamber further includes glycine in one of the embodiments, and the concentration of the glycine is
0.1mg/mL~50mg/mL.
The first chamber and/or the second chamber further include polysaccharide in one of the embodiments,.
The first chamber and/or the second chamber further include silica in one of the embodiments,.
First electron mediator and/or second electron mediator are selected from oxidation in one of the embodiments,
The electron mediator of state.
The electron mediator of the oxidation state is selected from the quinone of oxidation state in one of the embodiments, the quinone of oxidation state spreads out
One of biology, ferrocene, the iron cyanide and ruthenium salt are a variety of.
The pH value of first buffer and/or second buffer is 5~9 in one of the embodiments,.
Glutamic-oxalacetic transaminease Electrochemical Detection composition described in a kind of is used for glutamic-oxalacetic transaminease electrochemical sensor in preparation
Detection material in application.
A kind of glutamic-oxalacetic transaminease electrochemical sensor, the glutamic-oxalacetic transaminease electrochemical sensor include the anti-of multiple connections
Ying Chi, the reaction tank are provided with reaction reagent layer, and the reaction reagent layer of at least one reaction tank is by described
Glutamic-oxalacetic transaminease Electrochemical Detection composition in the first chamber formed through dry, at least one described reaction tank
The reaction reagent layer be by the second chamber in the glutamic-oxalacetic transaminease Electrochemical Detection composition through drying
It is formed.
A kind of detection method of glutamic-oxalacetic transaminease, using the glutamic-oxalacetic transaminease electrochemical sensor, and including following
Step:
Measure the first reaction reagent layer that the sample of known glutamic-oxalacetic transaminease concentration is formed in the first chamber
In the first current value and the second chamber formed the second reaction reagent layer in the second current value, by institute
It states the second current value and subtracts first current value and obtain relative current value；
The relative current value is measured according to default glutamic-oxalacetic transaminease concentration gradient, obtains glutamic-oxalacetic transaminease concentration and described
The standard curve of relative current value；
Measure the relative current value of sample to be tested；And
The relative current value of the sample to be tested is substituted into the standard curve, obtains the paddy in the sample to be tested
The concentration of careless transaminase.
The glutamic-oxalacetic transaminease Electrochemical Detection composition of the invention be used for detection sample in glutamic-oxalacetic transaminease into
The principle of row quantitative detection, the electrochemical quantitative detection of glutamic-oxalacetic transaminease turns ammonia in millet straw for ASPARTIC ACID and l-Alanine
Oxaloacetic acid is formed under the catalytic action of enzyme and glutamic acid, oxaloacetic acid form third under the catalytic action of oxaloacetic decarboxylase
Ketone acid and carbon dioxide, pyruvic acid under the action of pyruvate oxidase betatopic and be oxidized, electronics is caught by electron mediator
It obtains and exports in the form of electric current.The concentration of the pyruvic acid generated in detection sample can be characterized according to current value.If detection
There are endogenic pyruvic acid in sample, and the testing result of glutamic-oxalacetic transaminease can be made to increase compared with actual value, cause testing result
Inaccuracy.The pyruvate oxidase of the first chamber of the invention can carry out the endogenous pyruvic acid in detection sample
Detection, the second chamber can be to the endogenous pyruvic acid and glutamic-oxalacetic transaminease that detect in sample and oxaloacetic acid decarboxylations
The summation for the pyruvic acid that enzymatic ASPARTIC ACID and l-Alanine are formed is detected, the electricity that the second chamber is formed
The difference that stream subtracts the electric current that the first chamber is formed can characterize the concentration of the glutamic-oxalacetic transaminease in detection sample, thus
The interference of the endogenous pyruvic acid in different detection samples is excluded, testing result is more accurate.
Detailed description of the invention
Fig. 1 is the structural schematic diagram of the glutamic-oxalacetic transaminease electrochemical sensor of one embodiment of the invention；
Fig. 2 is the glutamic-oxalacetic transaminease concentration of one embodiment of the invention and the standard curve photo of current differential relationship.
Specific embodiment
In order to make the objectives, technical solutions, and advantages of the present invention clearer, by the following examples, it and combines attached
Figure, to glutamic-oxalacetic transaminease Electrochemical Detection composition of the invention, its application, electrochemical sensor and detection method.Carry out into
One step is described in detail.It should be appreciated that described herein, specific examples are only used to explain the present invention, is not used to limit this hair
It is bright.
The embodiment of the present invention provides a kind of glutamic-oxalacetic transaminease Electrochemical Detection composition, including first chamber and second group
Object is closed, the first chamber includes water, the first buffer, the first electrolyte, first surface activating agent, the first electron mediator
And pyruvate oxidase, the second chamber include water, the second buffer, the second electrolyte, second surface activating agent,
Two electron mediators, ASPARTIC ACID, l-Alanine, oxaloacetic decarboxylase and pyruvate oxidase.
The glutamic-oxalacetic transaminease Electrochemical Detection composition of the embodiment of the present invention is used to turn the millet straw in detection sample
Adnosine deaminase carries out quantitative detection, and the principle of the electrochemical quantitative detection of glutamic-oxalacetic transaminease is ASPARTIC ACID and l-Alanine in paddy
Oxaloacetic acid and glutamic acid are formed under the catalytic action of careless transaminase, oxaloacetic acid is under the catalytic action of oxaloacetic decarboxylase
Form pyruvic acid and carbon dioxide, pyruvic acid under the action of pyruvate oxidase betatopic and be oxidized, electronics is by electronics matchmaker
Mediator is captured and is exported in the form of electric current.The concentration of the pyruvic acid generated in detection sample can be characterized according to current value.Such as
There are endogenic pyruvic acid in fruit detection sample, and the testing result of glutamic-oxalacetic transaminease can be made to increase compared with actual value, cause to examine
Survey result inaccuracy.The pyruvate oxidase of the first chamber of the invention can be to the endogenous acetone in detection sample
Acid is detected, and the second chamber can be to the endogenous pyruvic acid and glutamic-oxalacetic transaminease and oxalyl second in detection sample
The summation for the pyruvic acid that acid decarboxylase catalysis ASPARTIC ACID and l-Alanine are formed is detected, the second chamber shape
At electric current subtract the difference of the electric current that the first chamber is formed and can characterize the dense of glutamic-oxalacetic transaminease in detection sample
Degree, to exclude the interference of the endogenous pyruvic acid in different detection samples, testing result is more accurate.
In one embodiment, first electron mediator is selected from the electron mediator of oxidation state.The electricity of the oxidation state
Sub- mediator captures the electronics during redox reaction, exports in the form of electric current.In one embodiment, the oxidation
The electron mediator of state can be selected from organic electronic mediator or inorganic electronic mediator, and the organic electronic mediator can select
One of the quinone of autoxidation state, the quinone derivative of oxidation state and ferrocene are a variety of, and the inorganic electronic mediator can select
From one of the iron cyanide and ruthenium salt or a variety of.Preferably, the electron mediator of the oxidation state can be the iron cyanide.?
In one embodiment, the concentration of first electron mediator in the second chamber can be 1mg/mL~50mg/mL.Institute
The type and concentration for stating the second electron mediator can be identical or different with first electron mediator.Preferably, described
The type and concentration of two electron mediators are identical as first electron mediator, guarantee the first chamber and described second
The electronic transmission performance for the redox reaction that composition mediates is identical, excludes accidental error.
In the first chamber, the pyruvate oxidase is used to will test the endogenous pyruvic acid oxygen in sample
Change, pyruvic acid is made to lose electronics, the electronics lost is exported in the form of electric current by first electron mediator, to characterize
Detect the relative concentration of the endogenous pyruvic acid in sample.In one embodiment, the pyruvic acid in the first chamber
The concentration of oxidizing ferment can be 0.1mg/mL~10mg/mL.
In the second chamber, the bottom of the ASPARTIC ACID and the l-Alanine as the first catalysis reaction
Object detects the glutamic-oxalacetic transaminease in sample as catalyst, is catalyzed the ASPARTIC ACID and the l-Alanine forms grass
Ethyl acetoacetic acid and glutamic acid.Substrate of the oxaloacetic acid as the second catalysis reaction, the oxaloacetic decarboxylase is as second
It is catalyzed the catalyst of reaction, the oxaloacetic acid is catalyzed and forms pyruvic acid, is characterized by the forming amount of the pyruvic acid to be detected
The concentration of glutamic-oxalacetic transaminease in sample.In one embodiment, the ASPARTIC ACID in the second chamber is dense
Degree can be 0.1mg/mL~10mg/mL, and the concentration of the l-Alanine can be 0.1mg/mL~10mg/mL.Due to described
ASPARTIC ACID and the l-Alanine react according to the ratio of 1:1, it is preferred that the ASPARTIC ACID and described
The concentration of l-Alanine is identical.In one embodiment, the concentration of the oxaloacetic decarboxylase can be 0.1mg/mL~10mg/
mL.The concentration of the oxaloacetic decarboxylase can be fitted according to the concentration of the ASPARTIC ACID and the l-Alanine
Answering property adjusts.
In the second chamber, the pyruvate oxidase is used to will test endogenous pyruvic acid and the institute in sample
It states glutamic-oxalacetic transaminease and is catalyzed the pyruvate oxidation to be formed, pyruvic acid is made to lose electronics, the electronics lost passes through second electronics
Mediator exports in the form of electric current, to characterize the endogenous pyruvic acid and glutamic-oxalacetic transaminease catalysis shape in detection sample
At pyruvic acid summation concentration.The summation for the pyruvic acid that the second chamber measures subtracts the first chamber and measures
Endogenous pyruvic acid obtain the concentration for the pyruvic acid that the glutamic-oxalacetic transaminease is catalyzed, the glutamic-oxalacetic transaminease is catalyzed to obtain
The concentration of pyruvic acid and the concentration of the glutamic-oxalacetic transaminease be positively correlated.That is, the obtained current value of the second chamber and
The difference for the current value that first chamber obtains and the concentration of the glutamic-oxalacetic transaminease in detection sample are positively correlated.It is real one
It applies in example, the concentration of the pyruvate oxidase in the second chamber is 0.1mg/mL~10mg/mL.
Preferably, the second chamber further includes glycine, in the reaction system formed due to the second chamber
Chemical reaction it is more, heat production is more, and the glycine is as protective agent, for guaranteeing the reactant in the second chamber
The thermal stability and pH stability of system, guarantee the high activity of various enzymes.The concentration of the glycine in the second chamber
It can be 0.1mg/mL~50mg/mL.
Surfactant is used to disperse each component of the first chamber and each component of the second chamber.One
In embodiment, the first surface activating agent can be selected from polyethylene glycol octyl phenyl enzyme, such as Triton X-100；Polyoxy second
Alkene sorbitan mono-laurate, such as polysorbas20；Chlolic acid derivatives class, such as 3- [(3- gallbladder amidopropyl) dimethyl ammonium
Base] -1- propane sulfonic acid salt, NaTDC；Brij-35, such as Brij35；And quaternary ammonium salt, such as cetyl three
One of methyl bromide ammonium is a variety of.Concentration of the first surface activating agent in the first chamber can be
0.1mg/ml~50mg/ml.The type and concentration of the second surface activating agent can be with the kinds of the first surface activating agent
Class and concentration are identical or different.
Buffer solution plays buffer function in the glutamic-oxalacetic transaminease Electrochemical Detection composition, for maintaining the paddy
The pH relative stability of careless transaminase Electrochemical Detection composition.First buffer solution can be selected from Tris, 4- ethoxy
One of piperazine ethanesulfonic acid (HEPES), TES and phosphate buffered saline solution (PBS) are a variety of.Preferably, first buffering
The pH value of solution can be 5~9, and in the range, so that first chamber is more stable, redox reaction progress is rapider,
It is more advantageous to the detection accuracy of endogenous pyruvic acid in blood.Preferably, it is 5~9 that first buffer solution, which is pH,
Tris buffer solution.Tris buffer solution will not interfere the electric signal in endogenous pyruvic acid detection process.It is real one
It applies in example, in the first chamber, the concentration of first buffer solution can be 0.1mg/ml~20mg/ml.It is described
Second buffer concentration, pH and type can be identical or different with first buffer.Preferably, first buffer and
The concentration of second buffer, pH and type are identical, guarantee the first chamber and second chamber detection environment
Consistency, reduce pyruvic acid detection accidental error.
Electrolyte is in the glutamic-oxalacetic transaminease Electrochemical Detection composition for conducting electronics and conduction.In an embodiment
In, first electrolyte can be selected from one of sodium chloride, potassium chloride and sodium sulphate or a variety of.Preferably, described first
Electrolyte is sodium chloride.In the first chamber, the concentration of first electrolyte can be 0.1mg/ml~10mg/
ml.In the concentration range, the electrochemical reaction rates of the pyruvate oxidase and endogenous pyruvic acid can be made to accelerate, and
And it is more stable to carry out electrochemical reaction, improves detection repeatability.The concentration and type of second electrolyte can with it is described
Second electrolyte is identical or different.Preferably, the concentration and type of second electrolyte and second electrolyte can phases
Together, the consistency for guaranteeing the first chamber and second chamber detection environment, reduces the accidental mistake of pyruvic acid detection
Difference.
Preferably, the first chamber further includes polysaccharide.Effect of the polysaccharide in the first chamber is to subtract
The rate of few negative dissolution, improves detection precision.In one embodiment, the polysaccharide can be selected from sodium alginate, trehalose, sweet
Reveal one of alcohol, sorbierite, sucrose, starch and cellulose or a variety of.Preferably, the polysaccharide includes at least cellulose.Institute
Cellulose is stated for stablizing blood constitutent, avoids the negative dissolution of blood.In the first chamber, the polysaccharide concentration can be with
For 0.1mg/ml~10mg/ml.Preferably, the first chamber and further include silica.The silica can be with
The polysaccharide cooperates jointly, forms a skeleton structure, reduces negative dissolution of the blood in the first chamber, is conducive to mention
High detection repeatability.In the first chamber, the concentration of the silica can be 0.1mg/ml~10mg/ml.Together
Reason, the second chamber may include polysaccharide.The second chamber may include silica.The second chamber with
The type and concentration of polysaccharide in the first chamber can just as or it is different.The second chamber is combined with described first
The concentration of silica in object can just as or it is different.The cellulose and the silica can by concussion, stirring,
The modes such as ultrasound are dispersed in the first chamber or the second chamber.
The embodiment of the present invention also provides the application of glutamic-oxalacetic transaminease Electrochemical Detection composition described in one kind, the millet straw
Transaminase Electrochemical Detection composition can prepare the detection material for glutamic-oxalacetic transaminease electrochemical sensor, such as by described
The dry reaction reagent layer for forming glutamic-oxalacetic transaminease electrochemical sensor of glutamic-oxalacetic transaminease Electrochemical Detection composition.Implement one
In example, the first chamber can be used for being formed the first reaction reagent layer of the first electrochemical sensor, first electrification
Sensor is learned to be used to detect the endogenous pyruvic acid in sample；The first chamber can be used for being formed the second electrochemical sensing
Second reaction reagent layer of device, the endogenous pyruvic acid and millet straw that second electrochemical sensor is used to detect in sample turn ammonia
The summation for the pyruvic acid that enzymatic is formed.First electrochemical sensor and second electrochemical sensor are used cooperatively,
Obtain the concentration of the glutamic-oxalacetic transaminease in detection sample.First electrochemical sensor and second electrochemical sensor can
To be separately provided or be integrated in an electrochemical sensor.Preferably, first electrochemical sensor and described second
Electrochemical sensor is integrated in an electrochemical sensor, and the measurement of glutamic-oxalacetic transaminease can be realized by being once loaded, can
To save the usage amount of detection sample, trace blood is realized.
Referring to Fig. 1, the embodiment of the present invention also provides a kind of glutamic-oxalacetic transaminease electrochemical sensor, the glutamic-oxalacetic transaminease
Electrochemical sensor includes the reaction tank 101 of multiple connections, and the reaction tank 101 is provided with reaction reagent layer (not shown), until
The reaction reagent layer of a rare reaction tank 101 is by the glutamic-oxalacetic transaminease Electrochemical Detection composition
The first chamber is formed through dry, and the reaction reagent layer of at least one reaction tank 101 is by the paddy
The second chamber in careless transaminase Electrochemical Detection composition is formed through dry.
The glutamic-oxalacetic transaminease electrochemical sensor uses multi-channel detection, and single injected sampling may be implemented, detect sample respectively
Endogenous pyruvic acid and glutamic-oxalacetic transaminease in this are catalyzed the sum of the pyruvic acid to be formed and endogenous pyruvic acid, to obtain millet straw
The concentration of transaminase.It realizes fewer detection sample (blood), accurately tests out the content of glutamic-oxalacetic transaminease, especially needle
To the middle-aged and the old, have an acupuncture treatment relatively difficult crowd, can obtain preferably detecting experience.
Specifically, the glutamic-oxalacetic transaminease electrochemical sensor may include sample introduction runner, the sample introduction runner be can wrap
Include sprue 102 and branch flow passage 103.The quantity of the branch flow passage 103 and the quantity of the reaction tank 101 are identical and mutual
It is corresponding.It detects sample to be added after the sprue, can enter along each reaction tank 101 along each branch flow passage 103, described the
The concentration of endogenous pyruvic acid is obtained in the first reaction tank 101 that one composition is formed, the formed in the second chamber
Obtain endogenous pyruvic acid in two reaction tanks 101 and glutamic-oxalacetic transaminease be catalyzed the pyruvic acid to be formed concentration summation.
Preferably, the branch flow passage 103 is two, and the reaction tank 101 is two, so that detection sample be avoided to flow into
Multiple reaction tanks 101 cause the waste of detection sample.
It is equipped with electrode layer 110 and reaction reagent layer in each reaction tank 101, and the reaction reagent layer in each reaction tank 12 is located at
On electrode layer 110.
Preferably, the glutamic-oxalacetic transaminease electrochemical sensor includes wall 200 and hydrophilic layer 300, the hydrophilic layer
300, wall 200 and the insulative substrate 100 are superposed from top to bottom.The hydrophilic layer 300, wall 200 and institute
The reaction tank 101 that recess can be surrounded between insulative substrate 100 is stated, the reaction reagent film can be set in the reaction tank
The surface of the insulative substrate 100 in 101.The wall 200 can be layers of two-sided.
Preferably, the hydrophilic layer 300 is equipped with the first venthole 310 being connected to the reaction tank 101.Implement at one
In example, label layer 400 is additionally provided on the hydrophilic layer 300.The label layer 400 is equipped with and is connected to first venthole 310
The second venthole 410.First venthole 310 and second venthole 410 are correspondingly arranged at the upper of the reaction tank 101
Side, and be connected to the reaction tank 101, so that it is logical to form siphon between the sprue 102 and the reaction tank 101
Road, using siphonic effect, sample solution can be directly sucked in from sample-adding end to the reaction tank 101 when being loaded end sample-adding
In, gas in former sprue 102, branch flow passage 103 and reaction tank 101 can be via first venthole 310 and described
The discharge of second venthole 410.
The embodiment of the present invention also provides a kind of detection method of glutamic-oxalacetic transaminease, using the glutamic-oxalacetic transaminease electrochemistry
Sensor, and the following steps are included:
Measure the first reaction reagent layer that the sample of known glutamic-oxalacetic transaminease concentration is formed in the first chamber
In the first current value and the second chamber formed the second reaction reagent layer in the second current value, by institute
It states the second current value and subtracts first current value and obtain relative current value；
The relative current value is measured according to default glutamic-oxalacetic transaminease concentration gradient, obtains glutamic-oxalacetic transaminease concentration and described
The standard curve of relative current value；
Measure the relative current value of sample to be tested；And
The relative current value of the sample to be tested is substituted into the standard curve, obtains the paddy in the sample to be tested
The concentration of careless transaminase.
Embodiment
First chamber and second chamber are configured according to Tables 1 and 2:
1 first chamber component of table
| Component | Content |
| Water | 1000ml |
| PH is the Tris-Hcl of 5-8 | 1.5g |
| Sodium chloride | 1g |
| Sodium alginate | 1g |
| PEG8000 | 1g |
| Cellulose | 1g |
| Silica | 1g |
| Triton X-100 | 2.5g |
| Pyruvate oxidase | 1g |
| The potassium ferricyanide | 2.5g |
2 second chamber component of table
The insulating substrate 100 for being provided with electrode is provided.Double branch flow passages 103 and two reactions are set on insulating substrate 100
Pond forms working electrode and reference electrode in reaction zone by the way of silk-screen printing.By configured first chamber and
Two compositions are put in differential responses area respectively, and working electrode is covered.First chamber and second chamber will be provided with
Insulating substrate 100 is dried at 50 DEG C, makes to be respectively formed the first reaction reagent layer and the second reaction reagent layer.
The electrode for being provided with the reaction reagent layer after drying is bonded with double-sided adhesive and hydrophilic film, is formed after steel rolling reality
Glutamic-oxalacetic transaminease electrochemical sensor test paper.Glutamic-oxalacetic transaminease electrochemical sensor test paper is cut with hobboing cutter or punching machine rushes
At each electrochemical sensor test paper having a size of 35mm × 6mm.
Glutamic-oxalacetic transaminease electrochemical sensor test paper is inserted into Auto Power On on the instrument of test, is fallen respectively in the suction of siphon
The content that glutamic-oxalacetic transaminease is added in place is 20g/L；100g/L；500g/L；The whole blood standard items of 1000g/l.
300mv voltage is selected, and is working electrode and is positive 300mv relative to the voltage of reference electrode, and in room temperature item
The content of glutamic-oxalacetic transaminease in sample is measured under part.The whole blood standard items of each concentration carry out 50 repeated experiments respectively, survey
The current value CV (coefficient of variation) tried out is within the scope of 1%-7%, and the testing mean of each concentration is as shown in table 3 below.
The current value of each concentration glutamic-oxalacetic transaminease of table 3
Referring to Fig. 2, the relationship of the glutamic-oxalacetic transaminease concentration and current differential measured according to table 3, obtains standard curve.Mark
Directrix curve equation are as follows: y=0.6607x+0.7999, wherein x represents the current differential of the second current value and the first current value, y generation
The concentration of table glutamic-oxalacetic transaminease.
In obtained standard curve, the deviation very little of the actual concentration of concentration and glutamic-oxalacetic transaminease that current value is converted into,
Illustrate that the test method of the present embodiment can accurately test out the content of glutamic-oxalacetic transaminease, and effectively eliminates the dry of pyruvic acid
It disturbs.
Each technical characteristic of embodiment described above can be combined arbitrarily, for simplicity of description, not to above-mentioned reality
It applies all possible combination of each technical characteristic in example to be all described, as long as however, the combination of these technical characteristics is not deposited
In contradiction, all should be considered as described in this specification.
The embodiments described above only express several embodiments of the present invention, and the description thereof is more specific and detailed, but simultaneously
Limitations on the scope of the patent of the present invention therefore cannot be interpreted as.It should be pointed out that for those of ordinary skill in the art
For, without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to guarantor of the invention
Protect range.Therefore, the scope of protection of the patent of the invention shall be subject to the appended claims.
Claims (14)
1. a kind of glutamic-oxalacetic transaminease Electrochemical Detection composition, which is characterized in that including first chamber and second chamber, institute
Stating first chamber includes water, the first buffer, the first electrolyte, first surface activating agent, the first electron mediator and third
Ketone acid oxidizing ferment, the second chamber include water, the second buffer, the second electrolyte, second surface activating agent, the second electronics
Mediator, ASPARTIC ACID, l-Alanine, oxaloacetic decarboxylase and pyruvate oxidase.
2. glutamic-oxalacetic transaminease Electrochemical Detection composition according to claim 1, which is characterized in that the first chamber
In the pyruvate oxidase concentration be 0.1mg/mL~10mg/mL.
3. glutamic-oxalacetic transaminease Electrochemical Detection composition according to claim 1, which is characterized in that the second chamber
In the pyruvate oxidase concentration be 0.1mg/mL~10mg/mL.
4. glutamic-oxalacetic transaminease Electrochemical Detection composition according to claim 1, which is characterized in that the second chamber
In the concentration of the ASPARTIC ACID be 0.1mg/mL~10mg/mL, the concentration of the l-Alanine be 0.1mg/mL~
10mg/mL。
5. glutamic-oxalacetic transaminease Electrochemical Detection composition according to claim 4, which is characterized in that the oxaloacetic acid is de-
The concentration of carboxylic acid is 0.1mg/mL~10mg/mL.
6. glutamic-oxalacetic transaminease Electrochemical Detection composition according to claim 1, which is characterized in that the second chamber
It further include glycine, the concentration of the glycine is 0.1mg/mL~50mg/mL.
7. glutamic-oxalacetic transaminease Electrochemical Detection composition according to claim 1-6, which is characterized in that described
One composition and/or the second chamber further include polysaccharide.
8. glutamic-oxalacetic transaminease Electrochemical Detection composition according to claim 1-6, which is characterized in that described
One composition and/or the second chamber further include silica.
9. glutamic-oxalacetic transaminease Electrochemical Detection composition according to claim 1-6, which is characterized in that described
One electron mediator and/or second electron mediator are selected from the electron mediator of oxidation state.
10. glutamic-oxalacetic transaminease Electrochemical Detection composition according to claim 9, which is characterized in that the oxidation state
Electron mediator is selected from one of the quinone of oxidation state, the quinone derivative of oxidation state, ferrocene, the iron cyanide and ruthenium salt or more
Kind.
11. -6,10 described in any item glutamic-oxalacetic transaminease Electrochemical Detection compositions according to claim 1, which is characterized in that institute
The pH value for stating the first buffer and/or second buffer is 5~9.
12. a kind of -11 described in any item glutamic-oxalacetic transaminease Electrochemical Detection compositions according to claim 1 are used for paddy in preparation
Application in the detection material of careless transaminase electrochemical sensor.
13. a kind of glutamic-oxalacetic transaminease electrochemical sensor, which is characterized in that the glutamic-oxalacetic transaminease electrochemical sensor includes more
The reaction tank of a connection, the reaction tank are provided with reaction reagent layer, the reaction reagent of at least one reaction tank
Layer is by the first chamber warp in the described in any item glutamic-oxalacetic transaminease Electrochemical Detection compositions of claim 1-11
Dry to be formed, the reaction reagent layer of at least one reaction tank is by the described in any item millet straws of claim 1-11
The second chamber in transaminase Electrochemical Detection composition is formed through dry.
14. a kind of detection method of glutamic-oxalacetic transaminease, which is characterized in that use glutamic-oxalacetic transaminease according to claim 13
Electrochemical sensor, and the following steps are included:
The sample of known glutamic-oxalacetic transaminease concentration is measured in the first reaction reagent layer that the first chamber is formed
First current value and the second current value in the second reaction reagent layer that the second chamber is formed, by described the
Two current values subtract first current value and obtain relative current value；
The relative current value is measured according to default glutamic-oxalacetic transaminease concentration gradient, obtains glutamic-oxalacetic transaminease concentration and described opposite
The standard curve of current value；
Measure the relative current value of sample to be tested；And
The relative current value of the sample to be tested is substituted into the standard curve, the millet straw obtained in the sample to be tested turns
The concentration of adnosine deaminase.
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