CN106841069A - A kind of glucose oxidase activity quick measuring reagent box and its detection method - Google Patents
A kind of glucose oxidase activity quick measuring reagent box and its detection method Download PDFInfo
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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
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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
- 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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- G01N2001/387—Other diluting or mixing processes mixing by blowing a gas, bubbling
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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
- G01N2021/3185—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry typically monochromatic or band-limited
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Abstract
A kind of glucose oxidase activity rapid assay methods, liquid enzyme liquid is suitably diluted with buffer solution；Absorption 3.0mL is added in cuvette by the measure reagent that 37 DEG C of water-baths are incubated, cuvette is put into spectrophotometer, returned to zero at wavelength 500nm, then to the enzyme liquid to be measured for adding 0.1mL to be incubated by 37 DEG C of water-baths in cuvette, add timing simultaneously in cuvette, and pressure-vaccum is allowed to well mixed immediately, light absorption value is recorded every 1min later, continuously record 10min.
Description
Technical field
A kind of assay kit and its detection method in the present invention relates to laboratory inspection field, and in particular to Yi Zhongyong
In the quick detection kit and its detection method for determining glucose oxidase activity.
Background technology
Glucose oxidase (Glucose oxidase, GOD), its systematic naming method is β-D-Glucose oxidoreducing enzyme
(EC1.1.3.4), it can be catalyzed β-D-Glucose high specificity and be reacted with oxygen, glucose is oxidized into gluconic acid
And hydrogen peroxide.The enzyme is distributed widely in animal, plant and microbial body.Nineteen twenty-eight Muller is first in aspergillus niger without thin
Glucose oxidase is found in born of the same parents' extract solution.Nineteen sixty Kusai etc., Pazurp and Swoboddas in 1964 are respectively from penicillin
With purifying glucose oxidase in aspergillus niger.Nineteen ninety-five Petruccioli etc. produces grape glycosyloxy with the mutant strain of penicillin
Change enzyme.China prepares purifying technique from the glucose oxidase that begins one's study in 1986, formally puts into production within 1998.1999
The Ministry of Agriculture is set to 12 kinds allows one of feed enzyme preparation additive for using.
The glycoxidative generation hydrogen peroxide (H of glucose oxidase catalysis grape2O2), H2O2Can under Catalyzed Synthesis By Peroxidase
By 4-AA and the pink quinone imines of oxidation of phenol condensation generation, fundamental reaction is as follows:
During measure, in the reaction of the first step, glucose substrate requirement versus glucose oxidizing ferment is definitely excessive, so as to urge
It is pseudo first order reaction to change reaction, and reaction rate is directly proportional to the enzyme amount for adding, and unrelated with concentration of substrate.The reaction mesh of second step
Be accurately to determine the first step to react discharged H2O2Amount, this single step reaction needs peroxidase with respect to H2O2Definitely
It is excessive, make the H of generation2O2It is rapid to participate in reaction, to reach the purpose of the real time measure first step reaction rate.Reaction solution color
The amount of the quinone imines that depth is produced to Catalyzed Synthesis By Peroxidase is directly proportional, and the growing amount of quinone imines and H2O2Growing amount is into just
Than H2O2Vigor of the growing amount then to glucose oxidase in reaction solution is directly proportional.
How by above-mentioned measuring principle by absorbing wavelength, buffer type, pH value, reaction time etc. carry out excellent
Change, it is that technical staff will solve to produce a kind of simple structure, glucose oxidase activity quick measuring reagent box easy to use
Problem certainly.
The content of the invention
The technical problem to be solved in the invention is to provide a kind of detection reagent of quick measure glucose oxidase activity
Box and its detection method, it is intended to solve the above problems.
In order to achieve the above object, the technical scheme is that：A kind of glucose oxidase activity rapid assay methods,
It is characterised in that it includes following steps；
Liquid enzyme liquid is centrifuged 10~20min by 1000~3000RCF, the supernatant for being obtained is fitted with buffer solution
Work as dilution；
1.0mL phosphate buffers are drawn, 37 DEG C of water-baths is placed in and is incubated 5~10min；
Enzyme liquids of the 1.0mL by appropriate dilution is drawn, 37 DEG C of water-baths is placed in and is incubated 5~10min；
Draw phosphate buffer 20.0mL, phenol solution 2.0mL, 4-AA solution 1.0mL, peroxide
Enzyme solutions 1.0mL and glucose substrate solution 6.0mL, amounts to 30.0mL and mixes as reagent is determined, and takes mixed liquor and is placed in right amount
37 DEG C of water-baths are incubated 5~10min；
A clean cuvette is taken, absorption 3.0mL is added in cuvette by the measure reagent that 37 DEG C of water-baths are incubated, by colorimetric
Cup is put into spectrophotometer, is returned to zero at wavelength 500nm, is incubated by 37 DEG C of water-baths then to 0.1mL is added in cuvette
Enzyme liquid to be measured, add timing simultaneously in cuvette, and pressure-vaccum is allowed to well mixed immediately, records extinction every 1min later
Value, continuously records 10min；
The vigor of glucose oxidase in enzyme dilution is calculated according to formula：
In formula, XD：The vigor of glucose oxidase in enzyme dilution, unit is U/mL；ΔA500：In certain hour
Solution extinction value changes at wavelength 500nm；Δt：In the extinction value changes corresponding reaction time, unit is min；1：Cuvette is thick
Degree, unit is cm；12.9：MM absorptivity of generation colour developing thing quinone imines, unit is Lmmol-1.cm-1；1/2：Often
1mol hydrogen peroxide generates the amount of quinone imines material；VAlways：Reaction mixture cumulative volume, unit is mL；：VEnzymeThe volume of enzyme solutions,
Unit is mL；
The enzyme activity of glucose oxidase sample is calculated according to formula, solid sample unit is every gram of unit of enzyme activity (U/
G), its computing formula is：Every milliliter of (U/mL) its computing formula of fluid sample unit unit of enzyme activity is：In formula, X：The enzyme activity of glucose oxidase sample, unit is U/g (or U/mL)；Df：Taken enzyme sample is complete
Portion is diluted to the corresponding buffer solution volume of suitable concn, and unit is mL；m：The solid enzyme sample quality for weighing, unit is g；V：Amount
The liquid enzymes sample volume for taking, unit is mL.
A kind of glucose oxidase activity quick measuring reagent box, the phase is characterised by, including R1, R2, R3, R4 and R5 examination
Agent, each reagent component is with proportioning：
R1：Buffer solution 100-200mmol/L；
R2：D/W 1000mmol/L；
R3：Peroxidase Solution 125KU/L；
R4：4-AA solution 4mg/mL (19.68mmol//L)；
R5：Phenol solution 1.5mg/mL (15.9mmol//L)；
PH of buffer 5.0-7.0,
Described buffer solution can be but be not limited to phosphate buffer, acetate buffer, Tris buffer solutions,
McIlvaine buffer solutions, its pH is 5.0-7.0.Kit also further contains nonionic surfactant, wherein nonionic table
Face activating agent can be TWEEN Series, TritonX nonionic surfactants, and consumption can be 0.01-1%, primarily serve increasing
Molten, clarification.
Brief description of the drawings
Fig. 1 is enzyme activity determination range of linearity regression straight line
Fig. 2 is enzyme activity determination range of linearity regression straight line
Specific embodiment
The present invention is further described with reference to embodiment.
Embodiment 1
A kind of glucose oxidase rapid quantitative detection reagent box in the present embodiment is five kinds of reagents, including R1, R2,
R3, R4 and R5 reagent, each reagent component is with proportioning：
R1 reagents：PH6.0 phosphate buffer 1s 00mmol/L；
R2 reagents：D/W 1000mmol/L；
R3 reagents：Peroxidase Solution 0.5mg/L；
R4 reagents：4-AA solution 4mg/mL；
R5 reagents：Phenol solution 1.5mg/mL；
After mentioned reagent dissolving, bottle is distributed into, is made the reagent of liquid five, directly mixing can be used in specific proportions.
Embodiment 2
Using glucose oxidase in the glucose oxidase rapid quantitative detection reagent box detection zymotic fluid in embodiment 1
Activity, concrete operation step is as follows：
1) preparation of sample
Zymotic fluid is centrifuged 10min by 2000RCF, the supernatant for being obtained is both then to take supernatant buffer solution
Suitably diluted；
2) 1.0mL phosphate buffers are drawn, 37 DEG C of water-bath insulation 5min are placed in；
3) enzyme liquids of the 1.0mL by appropriate dilution is drawn, 37 DEG C of water-bath insulation 5min are placed in；
4) phosphate buffer 20.0mL, phenol solution 2.0mL, 4-AA solution 1.0mL, peroxidating are drawn
Thing enzyme solutions 1.0mL and glucose substrate solution 6.0mL, amounts to 30.0mL and mixes as reagent is determined, and takes mixed liquor and puts in right amount
In 37 DEG C of water-bath insulation 5min；
5) a clean cuvette is taken, is drawn in 3.0mL measure reagent (being incubated by 37 DEG C of water-baths) addition cuvette, will compared
Color cup is put into spectrophotometer, is returned to zero at wavelength 500nm；
6) then to 0.1mL enzyme liquids to be measured (being incubated by 37 DEG C of water-baths) are added in cuvette, in addition cuvette simultaneously
Timing, and pressure-vaccum is allowed to well mixed immediately, records light absorption value every 1min later, continuously records 10min；According to formula
And formulaCalculate the activity of glucose oxidase.
Embodiment 3
Pure enzyme solutions glucose oxidase is detected using the glucose oxidase rapid quantitative detection reagent box in embodiment 1
Activity, concrete operation step is as follows：
1) sample preparation：Enzyme sample is weighed into two parts of sample, 0.001g is accurate to, is placed in 100mL volumetric flasks, added about
40mL phosphate buffers, in hertz oscilltor or constant temperature oscillator concussion 30min, or use magnetic stirrer 30min,
Taking-up phosphate buffer constant volume simultaneously shakes up, and takes centrifuge on the storage liquid for shaking up and 10min is centrifuged with 4000r/min, then
Supernatant is taken suitably to be diluted with phosphate buffer.Fluid sample can be directly diluted with phosphate buffer, be determined
Hold；
2) 1.0mL phosphate buffers are drawn, 37 DEG C of water-bath insulation 5min are placed in；
3) enzyme liquids of the 1.0mL by appropriate dilution is drawn, 37 DEG C of water-bath insulation 5min are placed in；
4) phosphate buffer 20.0mL, phenol solution 2.0mL, 4-AA solution 1.0mL, peroxidating are drawn
Thing enzyme solutions 1.0mL and glucose substrate solution 6.0mL, amounts to 30.0mL and mixes as reagent is determined, and takes mixed liquor and puts in right amount
In 37 DEG C of water-bath insulation 5min；
5) a clean cuvette is taken, is drawn in 3.0mL measure reagent (being incubated by 37 DEG C of water-baths) addition cuvette, will compared
Color cup is put into spectrophotometer, is returned to zero at wavelength 500nm；
6) then to 0.1mL enzyme liquids to be measured (being incubated by 37 DEG C of water-baths) are added in cuvette, in addition cuvette simultaneously
Timing, and pressure-vaccum is allowed to well mixed immediately, records light absorption value every 1min later, continuously records 10min；
7) solid sample unit is every gram of unit of enzyme activity (U/g), by formulaAnd formulaCalculated；
Every milliliter of fluid sample unit unit of enzyme activity (U/mL), by formulaAnd formulaCalculated.
Enzyme activity unit U is defined：Under the above-described reaction conditions, it is per minute make 1 μm of ol glucose be oxidized to gluconic acid and
H2O2Required enzyme amount, is defined as an enzyme activity unit.
Embodiment 4
The pure enzyme solutions of glucose oxidase are detected using the glucose oxidase rapid quantitative detection reagent box in embodiment 1
The activity of dilution is as a result as follows：
The pure enzyme preparation of glucose oxidase is dissolved in pH7.0 phosphate buffers, appropriate dilution obtains enzyme liquid A
(0.66U), is further carried out gradient dilution, according to the absorbance change of continuous Spectrophotometric Continuous Determination 1-10min.Often
Individual concentration is repeated twice, and the average value for taking each time point absorbance carries out linear regression analysis, straight using regression analysis gained
Line slope calculates enzyme activity.Result is as shown in table 1.
The light absorption value rate of change regression analysis of table 1 and inspection
From table 1 it follows that linearly dependent coefficient of the enzyme dilution under different concentration is higher, basic is in enzyme
Liquid concentration increases and reduces trend, and main cause is that, as enzyme activity increases, substrate is no longer definitely excessive for enzyme activity, reacts to one
Order reaction dynamics has deviateed.P value is respectively less than 0.01, therefore the change of light absorption value is sternly in 10min under each concentration
The linear change of lattice, therefore only change over time from absorbance and be difficult to judge the enzyme activity determination range of linearity, it is necessary to further analysis.
Enzyme activity is calculated using the slope of gained regression straight line, linear regression analysis is carried out to relative concentration with enzyme activity, and
Area Results mapping to relative enzyme liquid A concentration 0.1~0.5, is shown in Figure of description 1.
The enzyme activity determination range of linearity regression analysis of table 2 and inspection
From table 2 it can be seen that the concentration in relative enzyme liquid A is coefficient correlation and R in the range of 0.1~0.52Value is maximum,
Therefore enzyme activity determination value and concentration are in strict linear relationship in this concentration range.After the upper limit of concentration range increases, though
So there is the p value of very little, but from coefficient correlation, it is stricter from linear relationship in 0.1~0.5 concentration range, say
Bright book accompanying drawing 1 has also confirmed this point, thus may determine that 0.055~0.207U/mL is in the range of linearity of the assay method.
In practical application, enzyme liquid concentration can be easily judged with the size of extinction value changes per minute whether in linear scope,
Corresponding extinction value changes per minute are 0.011~0.042 as can be seen from Table 1.
It is cumbersome due to calculating Δ A/ Δs t with the method for linear regression, the practicality of continuous AAS is reduced,
Therefore attempt taking two-point method to calculate the point in the range of linearity.Choose respectively first group (2min and 6min), second group
(2min and 8min) and the 3rd group (2min and 10min) two groups of light absorption values are analyzed, as a result as shown in table 3, table 4 and table 5.
The two-point method of table 3 and standard method measurement result compare (first group)
The two-point method of table 4 and standard method measurement result compare (second group)
The two-point method of table 5 and standard method measurement result compare (the 3rd group)
By analysis, each concentration point in linear scope, take enzyme activity that the 3rd group of light absorption value two-point method obtains with
The enzyme activity that standard method is measured is fairly close, and maximum relative error is only 0.85%, second group of enzyme activity value relative error for measuring
Maximum is no more than 1.36%, can be with the measured value of alternate standard method.First group of enzyme activity value relative error for measuring most very much not surpasses
3.70% is crossed, it is for requiring that very not strict analysis can also receive and more quick.Therefore, for continuous point of standard
Light photometry, two-point method is more practical.
3 kinds of glucose oxidase samples of separate sources are chosen, 4000rpm/min centrifugations 10min, takes after appropriate dilution
Clear liquid determines enzyme activity.The light absorption value of 1~10min is recorded, record per minute is once.By formula (1) as can be seen that calculating
Enzyme activity is not need time point and the strict corresponding relation of light absorption value, it is only necessary to the light absorption value delta data of time period,
And Δ A/ Δs t is in the range of linearity that method is allowed.Light absorption value when taking 2~10min carries out linear regression analysis, while
Enzyme activity is calculated with standard method, and selects light absorption value during 2min and 10min to be calculated with two-point method, as shown in table 6.
The sample enzyme activity determination light absorption value rate of change regression analysis of table 6 is calculated with enzyme activity
Table 6 shows that sample enzyme activity determination light absorption value is good in 2~10min linear relationships, and standard method and two-point method
Result is highly consistent, directly can be determined using two-point method, and follow-up work can be carried out with this.
Embodiment 5
Pure enzyme solutions glucose oxidase is detected using the glucose oxidase rapid quantitative detection reagent box in embodiment 1
Activity, it is as a result as follows：
The pure enzyme preparation of glucose oxidase is dissolved in pH6.0 phosphate buffers, appropriate dilution obtains enzyme liquid enzyme liquid B
(0.59U), is further carried out gradient dilution, according to the absorbance change of continuous Spectrophotometric Continuous Determination 1-10min.Often
Individual concentration is repeated twice, and the average value for taking each time point absorbance carries out linear regression analysis, straight using regression analysis gained
Line slope calculates enzyme activity.Result is as shown in table 7.
The light absorption value rate of change regression analysis of table 7 and inspection
Linearly dependent coefficient under different concentration is higher, and basic is in increase with enzyme liquid concentration and reducing trend, main
It is that, as enzyme activity increases, substrate is no longer definitely excessive for enzyme activity, and reaction has deviateed first-order kinetics to want reason.p
Value is respectively less than 0.01, therefore the change of light absorption value is strict linear change in 10min under each concentration, therefore only from suction
Luminosity changes over time and is difficult to judge the enzyme activity determination range of linearity, it is necessary to further analysis.
Enzyme activity is calculated using the slope of gained regression curve, linear regression analysis is carried out to relative concentration with enzyme activity, and
Area Results mapping to relative enzyme liquid B concentration 0.1~0.5.
The enzyme activity determination range of linearity regression analysis of table 8 and inspection
As can be seen from Table 8, the concentration in relative enzyme liquid B is coefficient correlation and R in the range of 0.1~0.52Value is maximum,
Therefore enzyme activity determination value and concentration are in strict linear relationship in this concentration range.After the upper limit of concentration range increases, though
So there is the p value of very little, but from coefficient correlation, it is stricter from linear relationship in 0.1~0.5 concentration range, say
Bright book accompanying drawing 2 has also confirmed this point, thus may determine that 0.050~0.209U/mL is in the range of linearity of the assay method.
In practical application, enzyme liquid concentration can be easily judged with the size of extinction value changes per minute whether in linear scope,
The corresponding extinction value changes per minute of above range are about 0.010~0.042 as can be seen from Table 7.
It is cumbersome due to calculating Δ A/ Δs t with the method for linear regression, the practicality of continuous AAS is reduced,
Therefore attempt taking two-point method to calculate the point in the range of linearity.Choose respectively first group (2min and 6min), second group
(2min and 8min) and the 3rd group (2min and 10min) two groups of light absorption values are analyzed, as a result as shown in table 9, table 10 and table 11.
The two-point method of table 9 and standard method measurement result compare (first group)
The two-point method of table 10 and standard method measurement result compare (second group)
The two-point method of table 11 and standard method measurement result compare (the 3rd group)
By analysis, each concentration point in linear scope, take enzyme activity that second group of light absorption value two-point method obtains with
The enzyme activity that standard method is measured is fairly close, and maximum relative error is only 1.68%, the 3rd group of enzyme activity value relative error for measuring
Maximum is no more than 2.15%, can be with the measured value of alternate standard method.First group of enzyme activity value relative error for measuring most very much not surpasses
8.32% is crossed, it is for requiring that very not strict analysis can also receive and more quick.Therefore, for continuous point of standard
Light photometry, two-point method is more practical.
3 kinds of glucose oxidase samples of separate sources are chosen, 4000rpm/min centrifugations 10min, takes after appropriate dilution
Clear liquid determines enzyme activity.The light absorption value of 1~10min is recorded, record per minute is once.By formula as can be seen that calculating enzyme
Vigor is not need time point and the strict corresponding relation of light absorption value, it is only necessary to the light absorption value delta data of time period, and
Δ A/ Δs t is in the range of linearity that method is allowed.Light absorption value when taking 2~8min carries out linear regression analysis, while with
Standard method calculates enzyme activity, and selects light absorption value during 2min and 8min to be calculated, as shown in table 12.
The sample enzyme activity determination light absorption value rate of change regression analysis of table 12 is calculated with enzyme activity
Table 12 shows sample enzyme activity determination extinction value changes, good in 2~8min linear relationships, and standard method and two
Point method result is highly consistent, directly can be determined using two-point method.
Claims (3)
1. a kind of glucose oxidase activity rapid assay methods, it is characterised in that comprise the following steps：
1) liquid enzyme liquid is centrifuged 10~20min by 1000~3000RCF, the supernatant for being obtained is carried out suitably with buffer solution
Dilution；
2) 1.0mL phosphate buffers are drawn, 37 DEG C of water-baths is placed in and is incubated 5~10min；
3) enzyme liquids of the 1.0mL by appropriate dilution is drawn, 37 DEG C of water-baths is placed in and is incubated 5~10min；
4) phosphate buffer 20.0mL, phenol solution 2.0mL, 4-AA solution 1.0mL, peroxidase are drawn
Solution 1.0mL and glucose substrate solution 6.0mL, amounts to 30.0mL and mixes as reagent is determined, and takes mixed liquor and is placed in 37 in right amount
DEG C water-bath is incubated 5~10min；
5) a clean cuvette is taken, absorption 3.0mL is added in cuvette by the measure reagent that 37 DEG C of water-baths are incubated, by cuvette
It is put into spectrophotometer, is returned to zero at wavelength 500nm, is incubated by 37 DEG C of water-baths then to addition 0.1mL in cuvette
Enzyme liquid to be measured, adds timing simultaneously in cuvette, and pressure-vaccum is allowed to well mixed immediately, records light absorption value every 1min later,
Continuous record 10min；
6) vigor of glucose oxidase in enzyme dilution is calculated according to formula
In formula, XD：The vigor of glucose oxidase in enzyme dilution, unit is U/mL；ΔA500：In wavelength in certain hour
Solution extinction value changes at 500nm；Δt：In the extinction value changes corresponding reaction time, unit is min；1：Cuvette thickness, it is single
Position is cm；12.9：MM absorptivity of generation colour developing thing quinone imines, unit is L.mmol-1.cm-1；1/2：Per 1mol mistakes
Hydrogen oxide generates the amount of quinone imines material；VAlways：Reaction mixture cumulative volume, unit is mL；：VEnzymeThe volume of enzyme solutions, unit is
mL；
7) enzyme activity of glucose oxidase sample is calculated according to formula, solid sample unit is every gram of unit of enzyme activity (U/g),
Its computing formula is：Every milliliter of (U/mL) its computing formula of fluid sample unit unit of enzyme activity is：In formula, X：The enzyme activity of glucose oxidase sample, unit is U/g (or U/mL)；Df：Taken enzyme sample
The corresponding buffer solution volume of suitable concn all is diluted to, unit is mL；m：The solid enzyme sample quality for weighing, unit is g；V：
The liquid enzymes sample volume for measuring, unit is mL.
2. a kind of glucose oxidase activity quick measuring reagent box, the phase is characterised by, including R1, R2, R3, R4 and R5 reagent,
Each reagent component is with proportioning：
R1：Buffer solution 100-200mmol/L；
R2：D/W 1000mmol/L；
R3：Peroxidase Solution 125KU/L；
R4：4-AA solution 4mg/mL (19.68mmol//L)；
R5：Phenol solution 1.5mg/mL (15.9mmol//L)；
PH of buffer 5.0-7.0,
Described buffer solution can be but be not limited to phosphate buffer, acetate buffer, Tris buffer solutions, McIlvaine
Buffer solution, its pH is 5.0-7.0.
3. kit according to claim 2, the phase is characterised by that kit also further contains non-ionic surface active
Agent, wherein nonionic surfactant can be TWEEN Series, TritonX nonionic surfactants, and consumption can be 0.01-
1%, primarily serve solubilising, clarification.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107271393A (en) * | 2017-07-12 | 2017-10-20 | 广东岭南职业技术学院 | A kind of detection method of Fast Evaluation ferment fat-reducing effect |
| CN109280693A (en) * | 2018-11-12 | 2019-01-29 | 济南百斯杰生物工程有限公司 | A kind of method of Fast Evaluation glucose oxidase acid producing ability |
| WO2020257963A1 (en) * | 2019-06-24 | 2020-12-30 | 深圳迈瑞生物医疗电子股份有限公司 | Spreader and sample staining method |
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| CN109280693A (en) * | 2018-11-12 | 2019-01-29 | 济南百斯杰生物工程有限公司 | A kind of method of Fast Evaluation glucose oxidase acid producing ability |
| WO2020257963A1 (en) * | 2019-06-24 | 2020-12-30 | 深圳迈瑞生物医疗电子股份有限公司 | Spreader and sample staining method |
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