CN105665012B - A kind of Iron protein complexes Mimetic Peroxidase and its method for detecting hydrogen peroxide - Google Patents
A kind of Iron protein complexes Mimetic Peroxidase and its method for detecting hydrogen peroxide Download PDFInfo
- Publication number
- CN105665012B CN105665012B CN201610019876.XA CN201610019876A CN105665012B CN 105665012 B CN105665012 B CN 105665012B CN 201610019876 A CN201610019876 A CN 201610019876A CN 105665012 B CN105665012 B CN 105665012B
- Authority
- CN
- China
- Prior art keywords
- solution
- hydrogen peroxide
- iron
- albumen composition
- mimetic peroxidase
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J31/00—Catalysts comprising hydrides, coordination complexes or organic compounds
- B01J31/02—Catalysts comprising hydrides, coordination complexes or organic compounds containing organic compounds or metal hydrides
- B01J31/0234—Nitrogen-, phosphorus-, arsenic- or antimony-containing compounds
- B01J31/0235—Nitrogen containing compounds
-
- 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
Landscapes
- Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Engineering & Computer Science (AREA)
- Physics & Mathematics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Plasma & Fusion (AREA)
- Health & Medical Sciences (AREA)
- Materials Engineering (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The present invention relates to a kind of Iron protein complexes Mimetic Peroxidase, is the Iron protein complexes by ferric ion and albumen effect generation, has the function that similar natural peroxide enzyme.Hydrolyzed with this mimetic enzyme catalysis hydrogen peroxide and produce oxygen radical, hydrogen oxide donor dye makes generation colored dyes, using dye colour change and the absorption values in certain wave strong point, available for qualitative or quantitative detection hydrogen peroxide.
Description
Technical field
The present invention relates to a kind of iron-albumen composition Mimetic Peroxidase and its method for detecting hydrogen peroxide, belong to
The technical field of analytical chemistry.
Background technology
Hydrogen peroxide is commonly called as hydrogen peroxide, is a kind of important chemical products, in chemical industry, light industry, electronics, weaving, papermaking, dirt
The sector applications such as water process are extensive.It is each in food, environmental sanitation etc. because hydrogen peroxide has very strong oxidisability and Bleachability
Individual field, thus the content for detecting hydrogen peroxide in all kinds of samples has great importance.In field of food, hydrogen peroxide once by
Disinfectant and bleaching agent as food, too high residual peroxide can cause injury to alimentary tract to human body in food, cause
Canceration, inducing heart vessel disease.
The method of detection content of hydrogen peroxide has a variety of at present, mainly including stechiometry, electrochemical methods, fluorescence
Analytic approach and AAS etc..Horseradish peroxidase（HRP）It is a kind of peroxidase reagent being widely used.The enzyme
Containing positive iron-protoporphyrin, hydrogen peroxide oxidation hydrogen donor can be utilized, coloured oxidation type dye can be generated by reaction.Utilize
HRP catalysis characteristics, it has been widely used in the content for determining hydrogen peroxide in food or environmental sample.But HRP there is also
Limited source, purification difficult, it is expensive, not easy to maintain the shortcomings of, therefore, developing new analogue enztme has wide application
Prospect.
The content of the invention
It is an object of the invention to provide a kind of Mimetic Peroxidase and preparation method thereof, while present invention also offers
Utilize the method for analogue enztme detection hydrogen peroxide.
Mimetic Peroxidase of the present invention refers to that iron-albumen of ferric ion and protein effect generation is compound
Thing, this compound possess catalyzing hydrogen peroxide hydrolysis, have the function that similar natural peroxide enzyme.
The method of the present invention that content of hydrogen peroxide is determined using Mimetic Peroxidase, is referred to hydrogen peroxide
For substrate, using the Mimetic Peroxidase of above-mentioned preparation as catalyst, catalyzing hydrogen peroxide and hydrogen donor reaction, make hydrogen donor quilt
Oxidation generates coloured oxidation type dye, further detects hydrogen peroxide by the following method：（1）By observing dye colour
Change the presence of qualitative detection hydrogen peroxide；（2）Quantitatively detected by determining the absorption values of coloured oxidation dye
Aoxidize hydrogen content.
To achieve these goals, the present invention uses following technical scheme：
（One）The preparation of iron-albumen composition Mimetic Peroxidase：
Take protein solution and ferric ion solution to be added in the flask with constant temperature and agitating device, adjust the temperature to
Between 25-35 DEG C and keep constant, made with sodium hydroxide solution regulation solution ph between 8-11.Agitator is started, in perseverance
The lower strong stirring of temperature more than 24 hours.After question response, reaction solution is taken out, loads bag filter, using water as dialysis medium, dialysis
More than 36 hours, after eliminating unreacted free ferric ion, resulting solution was iron-albumen composition Mimetic Peroxidase
Solution.
In above-mentioned iron-albumen composition Mimetic Peroxidase preparation process, protein solution can use cow's serum
Albumin, human serum albumin, ovalbumin, livetin add water that protein solution is made, it is preferred to use bovine serum albumin(BSA).Egg
The concentration of white matter solution between 0.5% ~ 5%, is preferably controlled between 1% ~ 2% in control.Ferric ion solution can use chlorine
Change high ferro, ferric nitrate, ferric sulphate to be dissolved in water and be made, it is preferred to use ferric sesquichloride, which is dissolved in water, to be made.Ferric ion is molten
The concentration of liquid is controlled between 5 ~ 50 mmol/L, is preferably controlled between 20 ~ 30 mmol/L.Protein solution and high ferro from
The volume ratio of protein solution and ferric ion solution is controlled 10 in the mixed solution of son:1~2:Between 1, it is preferred to use 5:1
Ratio.
In above-mentioned iron-albumen composition Mimetic Peroxidase preparation process, the pH value of reaction temperature and mixed solution
It is the key factor in preparation process.Reaction temperature is too low, and reaction speed is slow, and required time is long；Reaction temperature is too high, can make
Protein is denatured, and causes reaction not carry out.Preferable reaction temperature is between 30 ± 2 DEG C and keeps constant.Mix molten
The regulation of the pH value of liquid should make the pH value of solution between 8 ~ 11, can according to different proteins isoelectric point difference and not
Together, its principle is to try to higher than isoelectric point but ferric ion can not be made to produce precipitation because alkalescence is excessive, and it is molten that it adjusts mixing
The method of liquid pH value is advisable using being slowly stirred down sodium hydroxide test solution is added dropwise dropwise not produce precipitation just to solution.Different eggs
The isoelectric point numerical value of white matter solution refers to as follows：Bovine serum albumin(BSA)（4.9）, human serum albumin（4.7-4.9）, ovalbumin
（4.71；4.59）, livetin（4.8-5.0）.
In the preparation process of above-mentioned iron-albumen composition Mimetic Peroxidase, the reaction time of mixed solution is 24
More than hour, preferably 36 hours；Dialysis time is more than 36 hours after reaction, preferably 72 hours.It should be checked after dialysis
Whether the ferric ion that dissociates in analysis liquid can detect, to ensure that dialysis is complete.
（Two）Utilize the method for iron-albumen composition Mimetic Peroxidase detection hydrogen peroxide：
Iron-albumen composition Mimetic Peroxidase solution is taken, sequentially adds hydrogen donor solution, hydrogen peroxide to be determined
Solution, add hydrochloric acid solution to adjust mixed liquor pH value to pH=2 or so, shake up, as need testing solution, replaced with method with equivalent water
Hydrogenperoxide steam generator to be determined makes blank solution.Need testing solution and blank solution are positioned in 37 DEG C of waters bath with thermostatic control and incubated
Educate 15 minutes, take out, hydrogen peroxide is detected according to following methods：（1）Compare need testing solution and blank solution reaction before and after color
Situation of change, if blank solution before the reaction after without significant change, need testing solution occur obvious color change afterwards before the reaction,
It can determine whether hydrogen peroxide in need testing solution be present, belong to qualitative detection；（2）If hydrogen peroxide in test sample be present, by test sample
Solution in 400-800nm wave-length coverages in determining absorption spectrum, according to the extinction number of degrees of maximum absorption wave strong point in absorption spectrum
Value seeks the concentration for calculating hydrogen peroxide, belongs to quantitative detection.
It is described in the method described above that hydrogen peroxide is detected using iron-albumen composition Mimetic Peroxidase
Hydrogen donor solution be 3,3 ', 5,5 '-tetramethyl biphenyl amide hydrochloride, the HCI solution of 3,3- diaminobenzidines four,
One kind in 4-AA solution, preferably TMB HCI solution.3,3’,5,5’-
The concentration of tetramethyl biphenyl amide hydrochloride is between 1 ~ 10mmol/L, preferably 5mmol/L, using dimethylformamide or
Dimethyl sulfoxide is prepared as solvent.
In the method described above that hydrogen peroxide is detected using iron-albumen composition Mimetic Peroxidase, iron-egg
The volume ratio of white compound Mimetic Peroxidase solution and hydrogen donor solution is 1:0.5~1:Between 2, preferably 1:1.Adopting
In by the use of TMB HCI solution as the method for hydrogen donor, color is by colourless after hydrogen donor is oxidized
It is changed into blueness, has absorption maximum near 657nm wavelength, using absorption values of the need testing solution at this wavelength and
Know that the absorption values of the reference substance solution of concentration compare, seek the concentration for calculating hydrogenperoxide steam generator in test sample.
As shown from the above technical solution, iron of the present invention-albumen composition Mimetic Peroxidase can be catalyzed
The reaction of hydrogen oxide and hydrogen donor, it is a kind of new Mimetic enzyme of peroxidase, available for the qualitative of hydrogen peroxide and quantifies
Detection.
The beneficial effects of the invention are as follows：Using cheap ferric ion and protein generation iron-albumen composition mistake
Oxide analogue enztme, preparation method is simple and easy to do, available for instead of expensive natural peroxide enzyme；The utilization invented
The method of iron-albumen composition analogue enztme detection hydrogen peroxide, mild condition, speed is fast, available for the qualitative of hydrogen peroxide and
Quantitative detection.
Brief description of the drawings
Fig. 1 is the absorption spectrum curve of iron-albumen composition Mimetic Peroxidase；
Fig. 2 is that iron-albumen composition Mimetic Peroxidase catalyzing hydrogen peroxide decomposes, oxidation 3,3 ', 5,5 '-tetramethyl
The extinction spectral curve of blue product obtained by benzidine；
Fig. 3 is to determine standard curve used in concentration of hydrogen peroxide using iron-albumen composition Mimetic Peroxidase, wherein
Abscissa is concentration of hydrogen peroxide, and ordinate is absorption values.
Embodiment
Beneficial effects of the present invention are made further explanation and description below by the drawings and specific embodiments.
Embodiment 1：The preparation of iron-albumen composition Mimetic Peroxidase
Iron(III) chloride hexahydrate 0.27g is taken in 50ml measuring bottles, be dissolved in water simultaneously constant volume, obtains ferric ion solution
（20mmol/L）.Take bovine serum albumin(BSA)（BSA）0.15g adds water 10ml to make dissolving in flask, and it is molten to add above-mentioned ferric ion
Liquid 2ml, pH value is adjusted to pH=10 with 10% sodium hydroxide solution under slow stirring.Flask is put into water bath with thermostatic control, installed
Agitating device, set bath temperature to 30 DEG C and to keep constant, react 36 hours under vigorous stirring.After completion of the reaction, incite somebody to action
Reaction solution takes out, and is fitted into the bag filter that molecular cut off is 8000, using 1000ml water as dialyzate, dialyses 72 hours.Saturating
During analysis, a dialyzate was changed every 12 hours.After dialysis, with phenol solution or other methods detection dialyzate
Whether ferric ion is remained, if remaining has ferric ion, continue to dialyse, if failing to detect ferric ion, dialysis is completed.
Solution in bag filter is taken out, is positioned in cillin bottle, it is stored refrigerated in refrigerator, produce iron-albumen composition peroxidating
Thing simulates enzyme solutions.Above-mentioned simulation enzyme solutions are taken, using water as blank, spectral scan, knot are carried out in 400 ~ 800nm wave-length coverages
Fruit is as shown in figure 1, as seen from Figure 1, iron-albumen composition Mimetic Peroxidase is without obvious UV absorption.
Embodiment 2：Color change and absorption spectrum during iron-albumen composition Mimetic Peroxidase detection hydrogen peroxide
The iron that the Example 1 is prepared-μ l of albumen composition Mimetic Peroxidase solution 50, sequentially adds 3,3 ',
5,5 '-tetramethyl biphenyl amide hydrochloride（5mmol/L）50 μ l, μ l of 0.3% hydrogenperoxide steam generator 50, hydrochloric acid solution（pH=2）
150 μ l, mix, as need testing solution.50 μ l of another water intaking replace hydrogenperoxide steam generator, and the amount of other materials is constant, ibid method
Prepare blank solution 1.Separately 50 μ l of water intaking replace iron-albumen composition Mimetic Peroxidase solution again, and the amount of other materials is not
Become, ibid method prepares blank solution 2.Need testing solution, blank solution 1, blank solution 2 are placed in 37 DEG C of waters bath with thermostatic control and are incubated
15 minutes, take out.Blank solution 1, blank solution 2 are near colourless, be incubated it is front and rear without significant change, after need testing solution is incubated
It is changed into blueness from colourless, can be with the presence of qualitative detection hydrogen peroxide by the change of need testing solution color., will using water as blank
Need testing solution is in carrying out spectral scan in 400 ~ 800nm wave-length coverages, as a result as shown in Figure 2.From Figure 2 it can be seen that iron-albumen is multiple
Compound Mimetic Peroxidase catalyzing hydrogen peroxide is decomposed so as to aoxidize the product of 3,3 ', 5,5 '-tetramethyl benzidine in 657nm
Wavelength at have absorption maximum, can utilize this wavelength at absorption values quantitative determination hydrogen peroxide concentration.
Embodiment 3：Utilize minimum detection limit during iron-albumen composition Mimetic Peroxidase qualitative detection hydrogen peroxide
It is determined that
Take 6 plastic centrifuge tubes, be separately added into 0.3% μ l of hydrogenperoxide steam generator 0,0.5 μ l, 1 μ l, 2 μ l, 5 μ l,
10 μ l, appropriate hydrochloric acid solution is separately added into each Guan Zhongzai（pH=2）It is 180 μ l to make liquor capacity, obtains 6 kinds of different peroxides
Change the need testing solution of hydrogen concentration.Iron-albumen composition mistake that embodiment 1 is prepared is added in 6 kinds of need testing solutions
Oxide simulation enzyme solutions 10 μ l, 3,3 ', 5,5 '-tetramethyl biphenyl amide hydrochloride（5mmol/L）10 μ l, mix, by 6
Centrifuge tube is placed in 37 DEG C of waters bath with thermostatic control and is incubated 15 minutes, takes out, and observes solution colour situation of change in centrifuge tube.As a result incubating
After the completion of educating, the test sample color of hydrogen peroxide is not added without significant change, and Color Color occurs in remaining 5 test sample pipe
Change, according to the volume of the hydrogenperoxide steam generator of addition, color is gradually deepened, by light blue to navy blue.Add amount of hydrogen peroxide
Solution is presented visually distinguishable light blue in minimum centrifuge tube, and lowest detection is used as using the concentration of hydrogen peroxide in this pipe
Limit, its concentration are 0.0008%.
Embodiment 4：Utilize the concentration of iron-albumen composition Mimetic Peroxidase quantitative determination hydrogen peroxide
（1）The preparation of standard curve：6 plastic centrifuge tubes are taken, respectively actual concentrations known to accurate addition（0.31%）Mistake
μ l of hydrogen peroxide solution 2,5 μ l, 8 μ l, 15 μ l, 25 μ l, 40 μ l, appropriate hydrochloric acid solution is separately added into each Guan Zhongzai（pH=
2）It is 360 μ l to make liquor capacity, obtains the standard liquid of 6 kinds of different concentration of hydrogen peroxide.Add in 6 kinds of standard liquids
Enter iron-μ l of albumen composition Mimetic Peroxidase solution 20,3,3 ', 5,5 '-tetramethyl biphenyl that embodiment 1 is prepared
Amide hydrochloride（5mmol/L）20 μ l, mix.6 centrifuge tubes are placed in 37 DEG C of waters bath with thermostatic control and are incubated 15 minutes, are taken out,
Absorption values are determined at 657nm wavelength.Using the concentration of hydrogen peroxide in standard liquid as abscissa, absorption values are vertical
Coordinate draws standard curve, and its result is as shown in Figure 3.
（2）Determination method：Plastic centrifuge tube is taken, adds μ l of hydrogenperoxide steam generator 20 to be determined, hydrochloric acid solution（pH=2）
340 μ l, then add embodiment 1 be prepared iron-μ l of albumen composition Mimetic Peroxidase solution 20,3,3 ', 5,
5 '-tetramethyl biphenyl amide hydrochloride（5mmol/L）20 μl.Centrifuge tube is placed in 37 DEG C of waters bath with thermostatic control and is incubated 15 minutes,
Take out, determine absorption values at 657nm wavelength immediately.Obtained absorption values are substituted into standard curve, obtained correspondingly
Hydrogen peroxide concentration, produce.
Claims (2)
- A kind of 1. iron-albumen composition Mimetic Peroxidase, it is characterised in that：It is by ferric ion and protein effect generation Iron-albumen composition, there is catalyzing hydrogen peroxide hydrolysis, similar natural peroxide enzyme；Itself another feature is that, It is to be prepared by following step：（1）Protein solution and ferric ion solution is taken to mix, adjusting the pH value of mixed solution makes Between 8-11,（2）Make the temperature of mixed solution between 25-35 DEG C and keep constant, strong stirring more than 24 hours,（3） Reaction product is loaded into bag filter, using water as dialysis medium, dialyses more than 36 hours, produces.
- 2. a kind of iron-albumen composition Mimetic Peroxidase as claimed in claim 1, the side of hydrogen peroxide is detected using it Method is as follows：Iron-albumen composition Mimetic Peroxidase solution is taken, it is molten to add TMB hydrochloride Liquid, hydrogenperoxide steam generator, with hydrochloric acid solution regulation mixed liquor pH value to pH=2 or so, shake up, be incubated in 37 DEG C of waters bath with thermostatic control 15 minutes, take out, front and rear color change situation is reacted according to reaction solution, whether qualitative judgement has hydrogen peroxide to participate in reaction；If Hydrogen peroxide be present and participate in reaction, by reaction solution in ultraviolet-visible spectrophotometer, surveyed in 400-800nm wave-length coverages Determine absorption spectrum, the concentration for calculating hydrogen peroxide is sought according to the absorption values of maximum absorption wave strong point in absorption spectrum.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610019876.XA CN105665012B (en) | 2016-01-13 | 2016-01-13 | A kind of Iron protein complexes Mimetic Peroxidase and its method for detecting hydrogen peroxide |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610019876.XA CN105665012B (en) | 2016-01-13 | 2016-01-13 | A kind of Iron protein complexes Mimetic Peroxidase and its method for detecting hydrogen peroxide |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105665012A CN105665012A (en) | 2016-06-15 |
| CN105665012B true CN105665012B (en) | 2018-01-26 |
Family
ID=56300393
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610019876.XA Expired - Fee Related CN105665012B (en) | 2016-01-13 | 2016-01-13 | A kind of Iron protein complexes Mimetic Peroxidase and its method for detecting hydrogen peroxide |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105665012B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110548017B (en) * | 2019-07-23 | 2021-09-21 | 中国科学院长春应用化学研究所 | Bovine serum albumin-iron nano-composite and preparation method and application thereof |
| CN112098402A (en) * | 2020-09-22 | 2020-12-18 | 程晓宏 | Method for rapidly detecting hydrogen peroxide based on peroxidase mimic enzyme activity |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101387606A (en) * | 2008-08-01 | 2009-03-18 | 中国科学院长春应用化学研究所 | Method for detecting hydrogen peroxide or glucose based on enzyme simulation by ferroferric oxide magnetic nanometer particle |
| CN103433484A (en) * | 2013-08-22 | 2013-12-11 | 福建医科大学 | Bovine serum albumin-platinum composite nanomaterial mimetic peroxidase |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5852564A (en) * | 1981-09-24 | 1983-03-28 | Amano Pharmaceut Co Ltd | Simple and quick microquantitative determination of hemoglobin |
-
2016
- 2016-01-13 CN CN201610019876.XA patent/CN105665012B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101387606A (en) * | 2008-08-01 | 2009-03-18 | 中国科学院长春应用化学研究所 | Method for detecting hydrogen peroxide or glucose based on enzyme simulation by ferroferric oxide magnetic nanometer particle |
| CN103433484A (en) * | 2013-08-22 | 2013-12-11 | 福建医科大学 | Bovine serum albumin-platinum composite nanomaterial mimetic peroxidase |
Non-Patent Citations (2)
| Title |
|---|
| Fe3O4 Magnetic Nanoparticles as Peroxidase Mimetics and Their Applications in H2O2 and Glucose Detection;Hui Wei et al.;《Anal. Chem.》;20080222;第80卷(第6期);第2250-2254页 * |
| 纳米材料模拟酶性质及其应用;梁敏敏 等;《东南大学学报(医学版)》;20110228;第30卷(第1期);第105页摘要 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105665012A (en) | 2016-06-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US3950226A (en) | Novel reagent and method for the determination of urea in biological fluids | |
| CN105021605B (en) | A kind of product and method for dissolved oxygen detection | |
| CN105665012B (en) | A kind of Iron protein complexes Mimetic Peroxidase and its method for detecting hydrogen peroxide | |
| CN107894509B (en) | A method of improving latex immunoturbidimetry antigen excess and the range of linearity | |
| CN102042979A (en) | Method for quickly detecting content of glucose in starch fermentation liquor | |
| CN105388150A (en) | Oxytetracycline test paper based on chromatic aberration comparison, using method and making method | |
| NO141910B (en) | PROCEDURE FOR REGULATING THE QUANTITY OF CHEMICALS IN LIQUIDS CONTAINED IN THE CELLULOSE INDUSTRY AND RELATED INDUSTRIES | |
| CN103512855B (en) | The detection method of reduced glutathione | |
| CN101887060A (en) | Kit for quantitatively detecting concentration of phenylalanine in blood of neonate by using fluorescence developing | |
| US8183051B2 (en) | Method and system for determining residual chlorine or chloramine concentration in solution by colorimetry | |
| CN108689933A (en) | A kind of hypochlorous fluorescence probe of quick high-selectivity analysis | |
| CN106093023A (en) | A kind of colorimetric sensor detecting mercury ion and preparation method thereof | |
| CN103954616A (en) | Method for detecting acetylcholine by simulating peroxidase based on bovine serum albumin-nano platinum | |
| CN107918019A (en) | A kind of detection method of fish anaphylactogen | |
| CN110514654A (en) | The rapid detection method of lactoperoxidase in a kind of cow's milk | |
| US4095948A (en) | Determination of uric acid | |
| CN108623522A (en) | A kind of hypochlorous method of quick high-selectivity detection | |
| CN110698390B (en) | Fluorescent probe for identifying bisulfite and preparation method and detection method thereof | |
| CN105717097A (en) | Sulfide ion detection kit based on bovine serum albumin-nano platinum/bismuth | |
| US3778384A (en) | Diagnostic composition for the quantitative determination of glucose | |
| CN113324931A (en) | Method for continuously and rapidly measuring ammonia nitrogen concentration in fresh water by using small system | |
| JP4940432B2 (en) | Analysis method of adenosine triphosphate contained in specimen | |
| CN105628700B (en) | Hydrogen peroxide detection kit based on bismuth-bovine serum albumin(BSA)-Platinum Nanoparticles | |
| CZ9900236A3 (en) | Method of qualitative and/or quantitative evaluation of a liquid and apparatus for making the same | |
| CN111323404B (en) | Fluorescence detection reagent, preparation method thereof and detection method of protamine or heparin |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20180126 Termination date: 20200113 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |