CN107101866A - A kind of glutathione reductase determines reagent and its preparation method and application - Google Patents
A kind of glutathione reductase determines reagent and its preparation method and application Download PDFInfo
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- CN107101866A CN107101866A CN201710365844.XA CN201710365844A CN107101866A CN 107101866 A CN107101866 A CN 107101866A CN 201710365844 A CN201710365844 A CN 201710365844A CN 107101866 A CN107101866 A CN 107101866A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/38—Diluting, dispersing or mixing samples
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/3103—Atomic absorption analysis
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
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Abstract
The present invention relates to biochemistry detection field, reagent and its preparation method and application is determined specifically, being related to a kind of glutathione reductase and determining reagent.The technical solution adopted by the present invention is：A kind of glutathione reductase determines reagent, it is characterised in that：The reagent includes reagent 1 and reagent 2, and the constituent of reagent 1 includes：Buffer solution, oxidized form of glutathione, stabilizer A；The constituent of reagent 2 includes：Buffer solution, reduced Coenzyme II, stabilizer B.It is an advantage of the invention that：Stabilization time of the glutathione reductase measure reagent provided by the present invention under 28 DEG C of use states of uncapping can reach that the stabilization time in 28 DEG C of closed lucifuge storing process can reach 10 15 months 20 30 days.
Description
Technical field
The present invention relates to biochemistry detection field, reagent is determined specifically, being related to a kind of glutathione reductase and determining reagent
And its preparation method and application.
Background technology
Glutathione reductase (glutathione reductase) is that one kind utilizes reduced-NAD (P) by oxidized form paddy
Enzyme of the sweet peptide of Guang (GS-SG) catalytic reaction into reduced form (GSH).
It can be seen that its reaction is irreversible in animal vegetable tissue, microorganism, yeast.
The enzyme of mouse liver almost can only be by the use of NADP as benefit enzyme, but the enzyme of the red blood cell in people also utilizes NAD to carry out
Effect.To cystine and homocystine without effect.Crystal can be produced from yeast.Seeming in plant tissue can be to dehydrogenase
Link with oxidizing ferment is worked.Glutathione reductase GS-SG, NADP and chloroplaset are mixed and use light irradiation, will
Oxygen is put, here it is Xi Er (Hill) reactions paid attention to.Contain flavine in this enzyme of Escherichia coli.
Glutathione is a kind of important cellular antioxidants.Missing glutathione reductase cell can be made to oxidant and
Antibiotic is more sensitive.
Traditional detection method adds dry powder using liquid is redissolved, and the existing molten existing use of reagent, stability is poor, is unsuitable for automatical analysis
Requirement, it is therefore necessary to developing a kind of new there is good stability and the glutathione reductase of detection accuracy to determine examination
Agent.
The content of the invention
The purpose of the present invention is to be lacking for the stability difference for overcoming traditional glutathione reductase assay method to exist
It is sunken that there is provided a kind of continual and steady glutathione under 2-8 DEG C of use state of uncapping and under 2-8 DEG C of closed lucifuge storage condition
Reduce enzymatic determination reagent.
Another object of the present invention be to provide preparation method that above-mentioned glutathione reductase determines reagent and
Application in measure.
To achieve the above object, the technical solution adopted by the present invention is：
A kind of glutathione reductase determines reagent, and the reagent includes reagent 1 and reagent 2, the composition of the reagent 1 into
Dividing includes：Buffer solution, oxidized form of glutathione, stabilizer A；The constituent of reagent 2 includes：Buffer solution, reduced Coenzyme II,
Stabilizer B；The buffer solution is phosphate buffer, TRIS buffer, glycine buffer, citric acid delay
Any of fliud flushing, carbonate buffer solution；The stabilizer A is in disodium ethylene diamine tetraacetate, magnesium chloride, potassium sorbate
One or more kinds of mixtures；The stabilizer B is one in sucrose, ascorbic acid, bovine serum albumin(BSA), potassium sorbate
Plant or two or more mixtures.
Wherein：
In the reagent 1, the concentration of oxidized form of glutathione is 0.1-1mmol/L；The final concentration of 50- of buffer solution
100mmol/L；Stabilizer A concentration is 1-2g/L.
In the reagent 2, the concentration of reduced Coenzyme II is 0.1-1g/L；The final concentration of 100-500mmol/ of buffer solution
L；Stabilizer B concentration is 1-10g/L.
Further technical scheme：
Also include calibration object, the constituent of the calibration object includes matrix, sterling and stabilizer C, and the matrix is slow
Mixture more than one or both of fliud flushing, casein, bovine serum albumin(BSA), the buffer solution be phosphate buffer,
Any of TRIS buffer, glycine buffer, citrate buffer solution, carbonate buffer solution；It is described
Sterling is glutathione reductase；The stabilizer C is one kind two in trehalose, ascorbic acid, disodium ethylene diamine tetraacetate
Plant the mixture of the above.
Wherein：
The concentration of sterling is 30-100U/L in the calibration object；Stabilizer C concentration is 1-10g/L；When the matrix
In when containing buffer solution, the concentration of buffer solution is 50-100mmol/L；When in the matrix contain casein when, casein it is dense
Spend for 1-10g/L；When containing bovine serum albumin(BSA) in the matrix, the concentration of bovine serum albumin(BSA) is 1-10g/L.
Further：
The pH of the reagent 1 is 7.0-8.0, and the pH of reagent 2 is 9.0-10.0.
The pH of the calibration object is 7.5-8.0.
A kind of glutathione reductase determines the preparation method of reagent, comprises the following steps：
A) preparation of reagent 1：The order by merging of each composition is not specially required in the preparation process of reagent 1, in room temperature bar
Under part, directly the buffer solution, oxidized form of glutathione, stabilizer A are carried out to mix simultaneously, in batches or step by step, as long as can obtain
To uniform stable phase, make mixing more uniform by stirring；
B) preparation of reagent 2：The preparation of reagent 2 is at ambient temperature, according to buffer solution, stabilizer B, reduced coenzyme
II order, is mixed successively, obtains uniform stable phase, makes mixing more uniform by stirring；
C) preparation of calibration object：The preparation of calibration object be at ambient temperature, according to matrix, stabilizer C, sterling it is suitable
Sequence, is mixed successively, obtains uniform stable phase, makes mixing more uniform by stirring.
A kind of glutathione reductase determines application of the reagent in glutathione reductase measure, comprises the following steps：
By sample to be tested, reagent 1, reagent 2 according to 6 μ L：150μL：50 μ L volume ratio (can according to the concentration of main component in reagent and
The difference of detecting instrument, carries out the adjustment of consumption in proportion) mix after stand 180-300 second, be subsequently placed in detecting instrument survey
2-3 minutes absorbances (A) are measured, absorbance change rate (△ A/min) per minute is calculated, wherein：37 DEG C of test temperature, detector
Device dominant wavelength 340nm, commplementary wave length 405nm, optical path 1cm carry out linear gauging using purified water and calibration object, measure sample to be tested
The concentration of Glutathione fabk polypeptide.
Wherein：
The sample to be tested be serum, blood plasma, calibration object, quality-control product or other contain solution of glutathione reductase,
The detecting instrument is the automatic clinical chemistry analyzer with double reagent function.
Due to the application of above-mentioned technical proposal, the present invention has the following advantages that compared with prior art：
1st, by above-mentioned technical proposal, glutathione reductase provided by the present invention, which determines reagent and uncapped at 2-8 DEG C, to be made
It can be reached 20-30 days with the stabilization time under state, the stabilization time in 2-8 DEG C of closed lucifuge storing process can reach
10-15 months.
2nd, the invention is easy to use, eliminating interference, obvious, sensitivity is high, pollution-free to biochemical instrument pipeline, makes it in paddy
There is important meaning in accurate, the quick measure of the sweet fabk polypeptide of Guang.
Embodiment
Technical scheme is further described with reference to specific embodiment, but the present invention is not limited to
Following embodiments.
Embodiment one
A kind of glutathione reductase determines reagent, and the reagent includes reagent 1 and reagent 2, the composition of the reagent 1 into
Dividing includes：Buffer solution, oxidized form of glutathione, stabilizer A, the buffer solution are phosphate buffer, and its is final concentration of
50mmol/L, the stabilizer A are disodium ethylene diamine tetraacetate, magnesium chloride, the mixture of potassium sorbate, wherein ethylenediamine tetrem
The final concentration of acid disodium, magnesium chloride and potassium sorbate is respectively 1.5g/L, the final concentration of 0.5mmol/L of oxidized form of glutathione；
The constituent of reagent 2 includes：Buffer solution, reduced Coenzyme II, stabilizer B, the buffer solution are trishydroxymethylaminomethane
Buffer solution, its final concentration of 250mmol/L, stabilizer B is sucrose, ascorbic acid, bovine serum albumin(BSA), the mixing of potassium sorbate
Thing, wherein sucrose, ascorbic acid, bovine serum albumin(BSA), the final concentration of potassium sorbate are respectively 5g/L, and reduced Coenzyme II is dense eventually
Spend for 0.5g/L；
The reagent also includes calibration object, and the constituent of the calibration object includes matrix, sterling and stabilizer C, the base
Matter is the mixture of buffer solution, casein and bovine serum albumin(BSA), and wherein buffer solution is TRIS buffer, its
Final concentration of 50mmol/L, the casein, the final concentration of bovine serum albumin are respectively 5g/L；The sterling be glutathione also
Protoenzyme, its final concentration of 80U/L；The stabilizer C is trehalose, the mixture of ascorbic acid, the trehalose, ascorbic acid
Final concentration is respectively 5g/L.
Above-mentioned glutathione reductase determines the preparation method of reagent, comprises the following steps：
A) preparation of reagent 1：The isometric purified water of dose volume is measured, from phosphate buffer, its is final concentration of
50mmol/L, is calculated by dose volume and weighs appropriate phosphate addition, regulation pH is 7.5, calculates and claims by dose volume
Appropriate oxidized form of glutathione is taken to add, final concentration of 0.5mmol/L；Calculated by dose volume and weigh appropriate second respectively
Edetate disodium, magnesium chloride, potassium sorbate are added, and final concentration is respectively 1.5g/L, is stirred, as reagent 1.
B) preparation of reagent 2：The isometric purified water of dose volume is measured, from TRIS buffer,
Final concentration of 250mmol/L, is calculated by dose volume and weighs appropriate trishydroxymethylaminomethane addition, regulation pH is 9.5,
Calculated by dose volume and weigh appropriate sucrose respectively, ascorbic acid, bovine serum albumin(BSA), potassium sorbate are added, final concentration
Respectively 5g/L, is calculated by dose volume and weighs appropriate reduced Coenzyme II addition, final concentration of 0.5g/L, stirring is equal
It is even, as reagent 2.
C) preparation of calibration object：The isometric purified water of dose volume is measured, from TRIS buffer,
Final concentration of 50mmol/L, is calculated by dose volume and weighs appropriate trishydroxymethylaminomethane addition, regulation pH is 7.5,
Calculated by dose volume and weigh appropriate casein respectively, bovine serum albumin(BSA) is added, final concentration is respectively 5g/L, by preparation
Volume is calculated and weighs appropriate trehalose respectively, ascorbic acid is added, and final concentration is respectively 5g/L, is calculated simultaneously by dose volume
Weigh appropriate glutathione reductase sterling to add, final concentration of 80U/L stirs, as calibration object.
Above-mentioned glutathione reductase determines application of the reagent in glutathione reductase measure, comprises the following steps：
By sample to be tested, reagent 1, reagent 2 according to 6 μ L：150μL：50 μ L volume ratio stands 240 seconds after mixing, and is subsequently placed in detection
2.5 minutes absorbances (A) are measured in instrument, absorbance change rate (△ A/min) per minute is calculated, wherein：Test temperature 37
DEG C, detecting instrument dominant wavelength 340nm, commplementary wave length 405nm, optical path 1cm carries out linear gauging using purified water and calibration object, presses
Formula：The concentration (U/L) of glutathione reductase=C standards * (△ A/minSample-△A/minBlank)/(△A/minStandard-△A/
minBlank) calculate sample Glutathione fabk polypeptide concentration, wherein the sample to be tested be serum, the detecting instrument for tool
There is the automatic clinical chemistry analyzer of double reagent function.
Embodiment two
A kind of glutathione reductase determines reagent, and the reagent includes reagent 1 and reagent 2, the composition of the reagent 1 into
Dividing includes：Buffer solution, oxidized form of glutathione, stabilizer A, the buffer solution are carbonate buffer solution, and its is final concentration of
80mmol/L, the stabilizer A are magnesium chloride, its final concentration of 1g/L, the final concentration of 1mmol/L of oxidized form of glutathione；Examination
The constituent of agent 2 includes：Buffer solution, reduced Coenzyme II, stabilizer B, the buffer solution are glycine buffer, and it is dense eventually
Spend for 500mmol/L, stabilizer B is sucrose, its final concentration of 1g/L, the final concentration of 1g/L of reduced Coenzyme II；
The reagent also includes calibration object, and the constituent of the calibration object includes matrix, sterling and stabilizer C, the base
Matter is the mixture of buffer solution and bovine serum albumin(BSA), and wherein buffer solution is TRIS buffer, its final concentration
For 80mmol/L, the final concentration of the bovine serum albumin is respectively 1g/L；The sterling is glutathione reductase, its final concentration
For 100U/L；The stabilizer C is trehalose, and the concentration of the trehalose is 10g/L.
Above-mentioned glutathione reductase determines the preparation method of reagent, comprises the following steps：
A) preparation of reagent 1：The isometric purified water of dose volume is measured, from carbonate buffer solution, its is final concentration of
80mmol/L, is calculated by dose volume and weighs appropriate carbonate addition, regulation pH is 7, calculates and weighs by dose volume
Appropriate oxidized form of glutathione is added, final concentration of 1mmol/L；Calculated by dose volume and weigh appropriate chlorination respectively
Magnesium, its final concentration is respectively 1g/L, is stirred, as reagent 1.
B) preparation of reagent 2：The isometric purified water of dose volume is measured, it is dense eventually from glycine buffer buffer solution
Spend for 500mmol/L, calculated by dose volume and weigh appropriate glycine buffer and add, regulation pH is 9, by dose volume
Calculate and weigh appropriate sucrose and add, its concentration is respectively 1g/L, calculated by dose volume and to weigh appropriate reduced form auxiliary
Enzyme II is added, final concentration of 1g/L, is stirred, as reagent 2.
C) preparation of calibration object：The isometric purified water of dose volume is measured, from TRIS buffer,
Final concentration of 80mmol/L, is calculated by dose volume and weighs appropriate trishydroxymethylaminomethane addition, regulation pH is 7, is pressed
Dose volume is calculated and weighs appropriate bovine serum albumin(BSA) and adds, and its final concentration of 1g/L is calculated and weighed by dose volume
Appropriate trehalose, final concentration of 10g/L is calculated by dose volume and is weighed appropriate glutathione reductase sterling addition,
Final concentration of 100U/L, stirs, as calibration object.
Above-mentioned glutathione reductase determines application of the reagent in glutathione reductase measure, comprises the following steps：
By sample to be tested, reagent 1, reagent 2 according to 6 μ L：150μL：50 μ L volume ratio stands 180 seconds after mixing, and is subsequently placed in detection
2 minutes absorbances (A) are measured in instrument, absorbance change rate (△ A/min) per minute is calculated, wherein：37 DEG C of test temperature,
Detecting instrument dominant wavelength 340nm, commplementary wave length 405nm, optical path 1cm carry out linear gauging, by public affairs using purified water and calibration object
Formula：The concentration (U/L) of glutathione reductase=C standards * (△ A/minSample-△A/minBlank)/(△A/minStandard-△A/
minBlank) calculate sample Glutathione fabk polypeptide concentration, wherein the sample to be tested be blood plasma, the detecting instrument for tool
There is the automatic clinical chemistry analyzer of double reagent function.
Embodiment three
A kind of glutathione reductase determines reagent, and the reagent includes reagent 1 and reagent 2, the composition of the reagent 1 into
Dividing includes：Buffer solution, oxidized form of glutathione, stabilizer A, the buffer solution are citrate buffer solution, and its is final concentration of
100mmol/L, the stabilizer A are the mixture of disodium ethylene diamine tetraacetate and potassium sorbate, wherein ethylenediamine tetra-acetic acid two
The final concentration of sodium and potassium sorbate is respectively 2g/L, the final concentration of 0.1mmol/L of oxidized form of glutathione；The composition of reagent 2 into
Dividing includes：Buffer solution, reduced Coenzyme II, stabilizer B, the buffer solution are citrate buffer solution, and its is final concentration of
100mmol/L, stabilizer B are the mixture of sucrose and potassium sorbate, and the final concentration of wherein sucrose and potassium sorbate is respectively
10g/L, the final concentration of 0.1g/L of reduced Coenzyme II；
The reagent also includes calibration object, and the constituent of the calibration object includes matrix, sterling and stabilizer C, the base
Matter is the mixture of casein and bovine serum albumin(BSA), the casein, the final concentration of bovine serum albumin be respectively 1g/L and
10g/L；The sterling is glutathione reductase, its final concentration of 30U/L；The stabilizer C is ascorbic acid, ethylenediamine tetraacetic
The mixture of acetic acid, the ascorbic acid, ethylenediamine tetra-acetic acid final concentration are respectively 1g/L.
Above-mentioned glutathione reductase determines the preparation method of reagent, comprises the following steps：
A) preparation of reagent 1：The isometric purified water of dose volume is measured, from citrate buffer solution buffer solution, its end
Concentration is 100mmol/L, is calculated by dose volume and weighs appropriate citric acid addition, regulation pH is 8, is calculated by dose volume
And weigh appropriate oxidized form of glutathione addition, final concentration of 0.1mmol/L；Calculate and weigh respectively appropriate by dose volume
Disodium ethylene diamine tetraacetate and potassium sorbate add, final concentration is respectively 2g/L, is stirred, as reagent 1.
B) preparation of reagent 2：The isometric purified water of dose volume is measured, it is dense eventually from citrate buffer solution buffer solution
Spend for 250mmol/L, calculated by dose volume and weigh appropriate citric acid and add, regulation pH is 10, is calculated by dose volume
And appropriate sucrose is weighed respectively and potassium sorbate is added, final concentration is respectively 10g/L, calculates and is weighed in right amount by dose volume
Reduced Coenzyme II add, final concentration of 0.1g/L stirs, as reagent 2.
C) preparation of calibration object：The isometric purified water of dose volume is measured, appropriate casein, bovine serum albumin is weighed
White to add, the casein, the final concentration of bovine serum albumin are respectively 1g/L and 10g/L, and regulation pH is 8, based on dose volume
Calculate and weigh appropriate ascorbic acid respectively and ethylenediamine tetra-acetic acid is added, final concentration is respectively 1g/L, is calculated by dose volume
And appropriate glutathione reductase sterling addition is weighed, final concentration of 30U/L stirs, as calibration object.
Above-mentioned glutathione reductase determines application of the reagent in glutathione reductase measure, comprises the following steps：
By sample to be tested, reagent 1, reagent 2 according to 6 μ L：150μL：50 μ L volume ratio stands 300 seconds after mixing, and is subsequently placed in detection
3 minutes absorbances (A) are measured in instrument, absorbance change rate (△ A/min) per minute is calculated, wherein：37 DEG C of test temperature,
Detecting instrument dominant wavelength 340nm, commplementary wave length 405nm, optical path 1cm carry out linear gauging, by public affairs using purified water and calibration object
Formula：The concentration (U/L) of glutathione reductase=C standards * (△ A/minSample-△A/minBlank)/(△A/minStandard-△A/
minBlank) calculate sample Glutathione fabk polypeptide concentration, wherein the sample to be tested be serum, the detecting instrument for tool
There is the automatic clinical chemistry analyzer of double reagent function.
Described above is preferred embodiment of the present utility model, it is noted that for the ordinary skill of the art
For personnel, on the premise of principle described in the utility model is not departed from, some improvement or replacement can also be made, these improvement
Or replace the protection domain that also should be regarded as invention.
Claims (10)
1. a kind of glutathione reductase determines reagent, it is characterised in that：The reagent includes reagent 1 and reagent 2, the reagent
1 constituent includes：Buffer solution, oxidized form of glutathione, stabilizer A；The constituent of reagent 2 includes：Buffer solution, reduction
Type codehydrogenase Ⅱ, stabilizer B；The buffer solution is phosphate buffer, TRIS buffer, glycine buffer
Any of liquid, citrate buffer solution, carbonate buffer solution；The stabilizer A be disodium ethylene diamine tetraacetate, magnesium chloride,
Mixture more than one or both of potassium sorbate；The stabilizer B is sucrose, ascorbic acid, bovine serum albumin(BSA), mountain
Mixture more than one or both of potassium sorbate.
2. a kind of glutathione reductase according to claim 1 determines reagent, it is characterised in that：In the reagent 1, oxygen
The concentration of change type glutathione is 0.1-1mmol/L；The final concentration of 50-100mmol/L of buffer solution；Stabilizer A concentration is
1-2g/L。
3. a kind of glutathione reductase according to claim 1 determines reagent, it is characterised in that：In the reagent 2, also
The concentration of prototype codehydrogenase Ⅱ is 0.1-1g/L；The final concentration of 100-500mmol/L of buffer solution；Stabilizer B concentration is 1-
10g/L。
4. a kind of glutathione reductase according to claim 1 or 2 or 3 determines reagent, it is characterised in that：Also include school
Quasi- product, the constituent of the calibration object includes matrix, sterling and stabilizer C, and the matrix is buffer solution, casein, ox blood
Mixture more than one or both of pure albumen；The buffer solution is phosphate buffer, trishydroxymethylaminomethane
Any of buffer solution, glycine buffer, citrate buffer solution, carbonate buffer solution；The sterling be glutathione also
Protoenzyme；The stabilizer C is a kind of two or more mixture in trehalose, ascorbic acid, disodium ethylene diamine tetraacetate.
5. a kind of glutathione reductase according to claim 4 determines reagent, it is characterised in that：In the calibration object
The concentration of sterling is 30-100U/L；Stabilizer C concentration is 1-10g/L；When containing buffer solution in the matrix, buffer solution
Concentration be 50-100mmol/L；When containing casein in the matrix, the concentration of casein is 1-10g/L；When the base
When containing bovine serum albumin(BSA) in matter, the concentration of bovine serum albumin(BSA) is 1-10g/L.
6. a kind of glutathione reductase according to claim 1 or 2 or 3 determines reagent, it is characterised in that：The reagent
1 pH is 7.0-8.0；The pH of reagent 2 is 9.0-10.0.
7. a kind of glutathione reductase according to claim 4 determines reagent, it is characterised in that：The pH of the calibration object
For 7.5-8.0.
8. a kind of glutathione reductase as described in any one in claim 1-7 determines the preparation method of reagent, it is special
Levy and be, comprise the following steps：
A) preparation of reagent 1：The order by merging of each composition is not specially required in the preparation process of reagent 1, at ambient temperature,
Directly the buffer solution, oxidized form of glutathione, stabilizer A are carried out to mix simultaneously, in batches or step by step, as long as can obtain uniform
Stable phase, by stirring make mixing more uniform；
B) preparation of reagent 2：The preparation of reagent 2 is at ambient temperature, according to buffer solution, stabilizer B, reduced Coenzyme II
Sequentially, mix successively, obtain uniform stable phase, make mixing more uniform by stirring；
C) preparation of calibration object：The preparation of calibration object be at ambient temperature, according to matrix, stabilizer C, sterling order, according to
Secondary mixing, obtains uniform stable phase, makes mixing more uniform by stirring.
9. a kind of glutathione reductase as described in any one in claim 1-7 determines reagent in glutathione reductase
Application in measure, it is characterised in that comprise the following steps：By sample to be tested, reagent 1, reagent 2 according to 6 μ L：150μL：50μL
Volume ratio mix after stand 180-300 second, be subsequently placed in detecting instrument 2-3 minutes absorbances (A) of measurement, every point of calculating
Clock absorbance change rate (△ A/min), wherein：37 DEG C of test temperature, detecting instrument dominant wavelength 340nm, commplementary wave length 405nm, light
Footpath 1cm, carries out linear gauging using purified water and calibration object, measures the concentration of sample to be tested Glutathione fabk polypeptide.
10. a kind of glutathione reductase according to claim 9 determines reagent in glutathione reductase measure
Using, it is characterised in that：The sample to be tested be serum, blood plasma, calibration object, quality-control product or other contain glutathione reductase
Solution, the detecting instrument be the automatic clinical chemistry analyzer with double reagent function.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110656155A (en) * | 2019-09-29 | 2020-01-07 | 江西乐成生物医疗有限公司 | Glutathione reductase determination reagent quality control product and preparation method thereof |
| CN110734952A (en) * | 2019-11-01 | 2020-01-31 | 江西乐成生物医疗有限公司 | Glutathione reductase detection kit and application |
| CN110747252A (en) * | 2019-09-29 | 2020-02-04 | 江西乐成生物医疗有限公司 | Glutathione reductase determination reagent calibrator and preparation method thereof |
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| CN110747252A (en) * | 2019-09-29 | 2020-02-04 | 江西乐成生物医疗有限公司 | Glutathione reductase determination reagent calibrator and preparation method thereof |
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