CN105295441B - A kind of stabilizer of color developing agent and its application - Google Patents
A kind of stabilizer of color developing agent and its application Download PDFInfo
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- CN105295441B CN105295441B CN201510811109.8A CN201510811109A CN105295441B CN 105295441 B CN105295441 B CN 105295441B CN 201510811109 A CN201510811109 A CN 201510811109A CN 105295441 B CN105295441 B CN 105295441B
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- stabilizer
- concentration
- agent
- color developing
- buffer
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- 125000004458 methylaminocarbonyl group Chemical group [H]N(C(*)=O)C([H])([H])[H] 0.000 description 1
- 239000007800 oxidant agent Substances 0.000 description 1
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- 150000002989 phenols Chemical class 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- XNGIFLGASWRNHJ-UHFFFAOYSA-L phthalate(2-) Chemical compound [O-]C(=O)C1=CC=CC=C1C([O-])=O XNGIFLGASWRNHJ-UHFFFAOYSA-L 0.000 description 1
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- Cosmetics (AREA)
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Abstract
The present invention relates to field of chemical detection, more particularly to a kind of stabilizer of color developing agent and its application.The present invention uses stabilizer of the azo dyes as color developing agent, has not only acted as good stabilization, and detection will not be interfered.Experiment shows, states the addition of azo dyes, improves the spontaneous colour developing of color developing agent and color developing agent is maintained more preferable coloration ability.In the check experiment of azo dyes is not added, coloring agent is all there occurs a degree of color change, and light absorption value is also there occurs obvious rise, and after using stabilizer provided by the invention, and dye color changes unobvious, and light absorption value change is not notable.Also, after high temperature is placed for a long time, remain to obtain good detection result.
Description
Technical field
The present invention relates to field of chemical detection, more particularly to a kind of stabilizer of color developing agent and its application.
Background technology
At present, to measure (qualitative, quantitative) method of target component, the redox reaction that enzyme participates in is applied mostly.
This method can usually be carried out according to following patterns：For example, generating oxidation material by target component to be measured, made using oxidizing ferment
The oxidation material is reacted with the color developing agent by aoxidizing generation color-developing compounds, the absorbance of colour developing caused by measure.The party
In method, the amount of color-developing compounds of the size of absorbance with being generated is suitable, the amount of the foregoing color-developing compounds generated and institute
The amount of the oxidation material of generation is suitable, and the amount of aforementioned oxidation material is suitable with the amount of target component.That is, by detecting
The colour developing (color-developing compounds generated) of generation, can by such redox reaction come measure indirectly target into
Point.
As described above, color developing agent is quite varied in clinical biochemical detection application.But the colour developing with redox active
Agent, it is often highly unstable.Usually occur before redox reaction artificial caused by adding enzyme starts, preserving
When produce nature colour developing it is such the problem of.Its reason be presumably due to：Such as in the case of foregoing wet system, in aforementioned solvents
Foregoing color developing agent becomes unstable, and in the case of responsibility, due to absorbing the moisture in air, thus foregoing color developing agent becomes not
Stablize etc..No matter in addition, wet system or responsibility, in ultraviolet when being preserved under the conditions of light irradiation, foregoing color developing agent also can be unstable
Fixedization and produce nature colour developing, this is also regarded as problem.
There is substantial amounts of method to be suggested and use to solve the above problems.
For example often used in the kit of liquid phase and freeze color developing agent (and other reagents), and be sealed.But
It is that such method requires complicated lyophilized technique step, and user is when in use, it is also desirable to add the operation of solution redissolution
Step.
Dry chemical strip has preferable stability, but its storage temperature is often room temperature, rather than kit is commonly used
Refrigerated storage temperature (2-8 DEG C).Therefore, to be equally that this area is urgently to be resolved hurrily ask the color developing agent stability problem in dry chemical strip
Topic.Substantial amounts of research has also been carried out to this both at home and abroad.
Such as a kind of method is：Using the surfactant and flavonoids system pigment of the alkyl with carbon number 8~16
Material.But this method formula is complicated, and reagent is being difficult to buy on the market, and protective agent in itself has test result certain again
Interference.
Also by make in liquid medium methylenum careuleum system color developing agent with carbon number be more than 12 alkyl season
Ammonium or its salt coexist, stabilizing it.But this method to methylenum careuleum system color developing agent for example：10- (Carboxymethylaminocarbonyl) -3,7-
Double (dimethylamino) phenthazine.Develop the color with preferable stability but to common Trinder ' s color developing agents and phenyl amines
Agent protectiveness is poor or without stabilization.
Also research uses Ar-NH-NH-CO-NH2Stable reagent color developing agent, or had using a kind of color stability agent
Reducing power more stronger than color developing agent, another color stability agent have between color developing agent and the first color stability agent
Reducing power.But the above method with the addition of stronger reducing substances, have necessarily to the redox reaction of color developing agent
Disturbing effect, in order to control interference in controlled range, to protectant with the more demanding of when concentration.Especially detecting
During analyte (creatinine, the transaminase) of low concentration, interference is notable.
In addition also have oxidation color former (color developing agent) and Fructoamino-acid-oxidase (FAOD) and peroxidase
At least one of (POD) coexist, stabilizing it.But this method can only stablize N- (carboxylic methylamino carbonyl) -4,4 '-two (two
Methylamino) diphenylamines sodium do not have to other color developing agents stability action or effect it is poor.
The above method all solves the problems, such as that color developing agent is unstable to varying degrees, but there are still it is different the defects of,
Therefore, further research is very necessary for the stabilizer of color developing agent.
The content of the invention
In view of this, the technical problem to be solved in the present invention is stabilizer and its application for providing a kind of color developing agent.This
The stabilizer that invention provides has good stablizing effect, even if remaining to play color developing agent at a higher temperature good steady
It is set for using, and the color developing effect of color developing agent is not interfered with.
The stabilizer of color developing agent provided by the invention, including azo dyes；
Azo dyes is selected from monoazo dyes, bisazo dye or polyazo dye；
Monoazo dyes is selected from tonyred l, tonyred 2, sudan orange B, the red, alizarin yellow R of temptation, orange G, lemon
Huang, amaranth, sunset yellow, tonyred G, Eriochrome Red B, acid red 18 or crocien orange G；
Bisazo dye is selected from acid red 66, Sudan black B, directly red 2B, alkaline brown g, tonyred B, tonyred 3, Soviet Union
Red red No. 4 or sudan red 7B；
Polyazo dye is direct black BN.
Azo dyes is kind and the most a class of dyes of quantity in current Dyestuff Market, because containing azo group in its molecule
And gain the name.At present, azo dyes is usually used in dyeing and the stamp of each fiber, and for leather, paper, soap, candle, wood
Material, straw, feather etc. dye and the coloring of paint, ink, plastics, rubber, food etc..The present invention is experimentally confirmed, even
Nitrogen dyestuff has stabilization to common color developing agent in biochemistry detection, can improve the spontaneous colour developing of color developing agent and make colour developing
Agent maintains more preferable coloration ability.Also, azo dyes can also reduce the interference to testing effect as stabilizer.
In certain embodiments, azo dyes is the composition of bisazo dye and polyazo dye, wherein, bisazo dye
The molar ratio of material and polyazo dye is 15:40.
In certain embodiments, azo dyes is the composition of monoazo dyes and bisazo dye, wherein, monoazo dye
The molar ratio of material and bisazo dye is 7：(4~20).
In an embodiment of the present invention, azo dyes be selected from crocien orange G, orange G, alkaline brown g, lemon yellow, alizarin yellow R,
Sudan orange B, Sudan black B, tonyred 3, tonyred 4, sudan red 7B, tonyred B, tonyred G, acid red 18, acid red
66th, Eriochrome Red B, the red, direct black BN of temptation or directly red 2B
In an embodiment of the present invention, azo dyes is selected from Sudan hong Ⅰ, tonyred 2, lures red, amaranth, the Sudan
Red G, Eriochrome Red B, acid red 18, acid red 66, directly red 2B, tonyred B, tonyred 3, tonyred 4 or tonyred
7B。
In an embodiment of the present invention, azo dyes be selected from tonyred 3, tonyred 4, sudan red 7B, tonyred B,
Tonyred G, acid red 18, acid red 66, directly Eriochrome Red B, red 2B or temptation are red.
In an embodiment of the present invention, the stabilizer of color developing agent further includes reducing agent, chelating agent, buffer, auxiliary and stablizes
Combination more than any of agent, surfactant or both.
Reducing substances refers to the chemical valence drop that the element that oxidation is played in oxidant can be reduced in chemical reaction
Low material.Experiment shows that, when stabilizer provided by the invention is used cooperatively with reducing substances, the stability of color developing agent can
Further to improve.
In an embodiment of the present invention, the concentration of reducing agent is 10 μm of ol/L~100 μm ol/L；
Reducing agent be selected from 1- Carbaphens, 1,5- diphenyl ureas, N, N, N- trimethylamino ureas, 4- Carbaphens,
Bisphenol-A, gallic acid, carbohydrazide, paracetamol or gentianic acid.
In certain embodiments, reducing agent is 1- Carbaphens, 4- Carbaphens, carbohydrazide or acetparaminosalol
Phenol.
In an embodiment of the present invention, the concentration of chelating agent is 20 μm of ol/L~150 μm ol/L；
Chelating agent be selected from ethylenediamine tetra-acetic acid, diethylene-triamine pentaacetic acid, anti-form-1,2- diaminocyclohexane-N, N,
N ', N '-tetraacethyl, O, O '-two (2- aminoethyls) ethylene glycol-N, N, N ', N '-tetraacethyl or nitrilotriacetic acid.
In certain embodiments, chelating agent EDTA, GEDTA or DTPA.
In an embodiment of the present invention, the concentration of buffer is 0.1mol/L；
Buffer is selected from citric acid buffer agent, phosphate buffer, acetate buffer agent or Good ' s buffers.
In certain embodiments, buffer is Hepes buffer solutions, phosphate buffer, MOPS buffer solutions or MES bufferings
Liquid.
In an embodiment of the present invention, the mass fraction of surfactant is 0.5%~1%；
The surfactant is selected from Tween series, Triton series, Brij series or Pluronic series.
In certain embodiments, surfactant TritonX100, Pluronic F-68 or Tween 20.
In an embodiment of the present invention, the mass fraction of auxiliary stabilizer is 1%~2%；
Auxiliary stabilizer is selected from carbohydrate or protide.
In certain embodiments, carbohydrate is sucrose, trehalose or sorbierite；Albumen is BSA.
In an embodiment of the present invention, color developing agent is selected from phenyl amines, phenols, Trinder, connection nitrogen class or imidazoles.
In certain embodiments, phenyl amines color developing agent is selected from TOPS (N- ethyls-N- (3- sulfopropyls) -3- methylaniline sodium
Salt), MAOS (N- ethyls-N- (2- hydroxyl -3- sulfopropyls) -3-5- dimethoxyanilines), MADB [N, N- double (4- sulphur butyls) -
3-5- dimethylanilines], TMB (3,3',5,5'-tetramethylbenzidine) or TOOS [N- ethyls-N- (2- hydroxyl -3- sulfopropyls) -
3- methylanilines sodium salt].
In certain embodiments, Trinder color developing agents are selected from 4AAP (4-AA), TBHBA (2,4,6- tri-
Bromo- 3- hydroxybenzoic acids) or DHBS (sodium 3,5-dichloro-2-hydroxybenzenesulfonate).
In certain embodiments, join nitrogen class color developing agent for ABTS [(2,2 '-azo-bis- (3- ethyl benzo thiazole phenanthroline -6- sulphurs
Acid) di-ammonium salts)].
In certain embodiments, color developing agent is selected from TMB, 4AAP, MAOS, MADB, DHBS or TBHBA.
In one embodiment of the invention, the stabilizer of coloring agent includes azo dyes, surfactant and buffer,
Wherein the concentration of azo dyes is 1 μm of ol/L~110 μm ol/L；The mass fraction of surfactant is 1%；The concentration of buffer solution
For 0.1mol/L.
Preferably, azo dyes is selected from sudan orange B, alizarin yellow R, crocien orange G, orange G, sunset yellow or lemon yellow；Table
Face activating agent is TritonX100；Buffer solution is MOPS buffer solutions.
Preferably, the stabilizer of coloring agent includes azo dyes of the concentration for 1 μm of ol/L, mass fraction is 1% table
Face activating agent, concentration are the buffer solution of 0.1mol/L.
Preferably, the stabilizer of coloring agent includes sudan orange B of the concentration for 1 μm of ol/L, and mass fraction is 1%
TritonX100, concentration are the MOPS buffer solutions of 0.1mol/L.
Preferably, the stabilizer of coloring agent includes azo dyes of the concentration for 110 μm of ol/L, mass fraction is 1%
Surfactant, concentration are the buffer solution of 0.1mol/L.
Preferably, the stabilizer of coloring agent includes alizarin yellow R of the concentration for 10 μm of ol/L, and concentration is the Tibetan of 100 μm of ol/L
Spend orange G, the TritonX100 that mass fraction is 1%, concentration is the MOPS buffer solutions of 0.1mol/L.
Preferably, the stabilizer of coloring agent includes azo dyes of the concentration for 25 μm of ol/L, mass fraction is 1% table
Face activating agent, concentration are the buffer solution of 0.1mol/L.
Preferably, the stabilizer of coloring agent includes orange G of the concentration for 5 μm of ol/L, concentration for 20 μm of ol/L sunset yellow,
Mass fraction is 1% TritonX100, and concentration is the MOPS buffer solutions of 0.1mol/L.
Preferably, the stabilizer of coloring agent includes azo dyes of the concentration for 5 μm of ol/L, mass fraction is 1% table
Face activating agent, concentration are the buffer solution of 0.1mol/L.
Preferably, the stabilizer of coloring agent includes lemon yellow of the concentration for 5 μm of ol/L, and mass fraction is 1%
TritonX100, concentration are the MOPS buffer solutions of 0.1mol/L.
Preferably, the stabilizer provided in this embodiment is used for the stabilization of coloring agent TMB.
In this embodiment, the molar ratio of azo dyes and coloring agent TMB are 1:10000、110:10000、25:10000、
Or 5:10000.
In this embodiment, the suitable services pH value of the stabilizer of coloring agent is 7.4.
In one embodiment of the invention, the stabilizer of coloring agent includes azo dyes and buffer, and wherein azo contaminates
The concentration of material is 0.1 μm of ol/L~1mmol/L；The concentration of buffer solution is 0.1mol/L.
Preferably, azo dyes is selected from acid red 66, Sudan black B, alkaline brown g, direct black BN, tonyred B or the Sudan
Red 7B；Buffer solution is phosphate buffer.
Preferably, the stabilizer of coloring agent includes azo dyes of the concentration for 0.1 μm of ol/L, concentration is 0.1mol/L's
Buffer solution.
Preferably, the stabilizer of coloring agent includes acid red 66 of the concentration for 0.1 μm of ol/L, and concentration is the phosphorus of 0.1mol/L
Phthalate buffer.
Preferably, the stabilizer of coloring agent includes azo dyes of the concentration for 50 μm of ol/L, concentration is 0.1mol/L's
Buffer solution.
Preferably, the stabilizer of coloring agent includes Sudan black B of the concentration for 50 μm of ol/L, and concentration is the phosphoric acid of 0.1mol/L
Salt buffer.
Preferably, the stabilizer of coloring agent includes azo dyes of the concentration for 55 μm of ol/L, concentration is 0.1mol/L's
Buffer solution.
Preferably, the stabilizer of coloring agent includes alkaline brown g of the concentration for 15 μm of ol/L, and concentration is direct for 40 μm of ol/L's
Black BN, concentration are the phosphate buffer of 0.1mol/L.
Preferably, the stabilizer of coloring agent includes azo dyes of the concentration for 1075 μm of ol/L, concentration 0.1mol/L
Buffer solution.
Preferably, the stabilizer of coloring agent includes tonyred B of the concentration for 15 μm of ol/L, and concentration is the Soviet Union of 1000 μm of ol/L
Red red 7B, concentration are the phosphate buffer of 0.1mol/L.
Preferably, the stabilizer provided in this embodiment is used for the stabilization of coloring agent 4AAP and MAOS.
In this embodiment, the ratio between the sum of molal quantity of the molal quantity of azo dyes and coloring agent 4AAP and MAOS is 0.1:
20000、50:20000、55:20000 or 1075:20000.
In this embodiment, the suitable services pH value of the stabilizer of coloring agent is 6.5.
In one embodiment of the invention, the stabilizer of coloring agent include azo dyes, reducing agent, auxiliary stabilizer and
The concentration of buffer, wherein azo dyes is 10 μm of ol/L~15 μm ol/L, and the concentration of reducing agent is the μ of 50 μm of ol/L~100
Mol/L, the mass fraction of auxiliary stabilizer is 2%, and the concentration of buffer is 0.1mol/L.
Preferably, azo dyes is red selected from direct red 2B, temptation；Reducing agent is 4- Carbaphens；Auxiliary stabilizer
For sucrose；Buffer solution is HEPES buffer solution.
Preferably, the stabilizer of coloring agent includes direct red 2B of the concentration for 10 μm of ol/L；Concentration is 50 μm of ol/L
4- Carbaphens；Mass fraction is 2% sucrose；Concentration is the HEPES buffer solution of 0.1mol/L.
Preferably, azo dyes be selected from for direct red 2B, temptation it is red；Reducing agent is 4- Carbaphens；Auxiliary is stablized
Agent is sucrose；Buffer solution is HEPES buffer solution.
Preferably, the stabilizer of coloring agent is red including the temptation that concentration is 15 μm of ol/L；Concentration is the 4- of 100 μm of ol/L
Carbaphen；Mass fraction is 2% sucrose；Concentration is the HEPES buffer solution of 0.1mol/L.
Preferably, the stabilizer provided in this embodiment is used for the stabilization of coloring agent 4AAP and MADB.
In this embodiment, the ratio of the molal quantity of azo dyes and the sum of the molal quantity of coloring agent 4AAP and MADB is 10:
20000 or 15:20000.
In this embodiment, the suitable services pH value of the stabilizer of coloring agent is 7.5.
In one embodiment of the invention, the stabilizer of coloring agent include azo dyes, chelating agent, auxiliary stabilizer and
Buffer, wherein, the concentration of azo dyes is 20 μm of ol/L, the concentration of chelating agent is 40 μm of ol/L, the quality of auxiliary stabilizer
Fraction is 2%, and the concentration of buffer is 0.1mol/L.
Preferably, azo dyes is tonyred 3；Chelating agent is EDTA；Auxiliary stabilizer is sucrose；Buffer is
HEPES。
Preferably, dying-stable agent includes tonyred 3 of the concentration for 20 μm of ol/L；Concentration is the EDTA of 40 μm of ol/L；
Mass fraction is 2% sucrose；Concentration is the HEPES of 0.1mol/L.
Preferably, the stabilizer provided in this embodiment is used for the stabilization of coloring agent 4AAP and MADB.
In this embodiment, the ratio of the molal quantity of azo dyes and the sum of the molal quantity of coloring agent 4AAP and MADB is 20:
20000。
In this embodiment, the suitable services pH value of the stabilizer of coloring agent is 7.5.
In one embodiment of the invention, the stabilizer of coloring agent include azo dyes, chelating agent, auxiliary stabilizer and
Buffer, wherein, the concentration of azo dyes is 35 μm of ol/L, the mass fraction of auxiliary stabilizer is 2%, the concentration of buffer is
0.1mol/L。
Preferably, azo dyes is tonyred 4；Auxiliary stabilizer is sucrose；Buffer is HEPES.
Preferably, dying-stable agent includes tonyred 4 of the concentration for 35 μm of ol/L；Mass fraction is 2% sucrose；It is dense
Spend the HEPES buffer solution for 0.1mol/L.
Preferably, the stabilizer provided in this embodiment is used for the stabilization of coloring agent 4AAP and MADB.
In this embodiment, the ratio of the molal quantity of azo dyes and the sum of the molal quantity of coloring agent 4AAP and MADB is 35:
20000。
In this embodiment, the suitable services pH value of the stabilizer of coloring agent is 7.5.
In one embodiment of the invention, the stabilizer of coloring agent includes azo dyes, reducing agent, chelating agent, surface
Activating agent, auxiliary stabilizer and buffer；Wherein, the concentration of azo dyes for 55 μm of ol/L~100 μm ol/L, reducing agent it is dense
The concentration spent for 10 μm of ol/L~50 μm ol/L, chelating agent is 100 μm of ol/L~140 μm ol/L, the quality of surfactant point
Number is 0.5%, the mass fraction of auxiliary stabilizer is 1%, and the concentration of buffer is 0.1mol/L.
Preferably, azo dyes is selected from direct red 2B, tonyred 3 or tonyred G；Reducing agent is carbohydrazide；Chelating
Agent is DTPA；Surfactant is Pluronic F-68；Auxiliary stabilizer is sorbierite；Buffer solution is MES buffer solutions.
Preferably, the stabilizer of color developing agent includes direct red 2B of the concentration for 100 μm of ol/L, and concentration is the carbon of 10 μm of ol/L
Hydrazides, concentration are the DTPA of 100 μm of ol/L, and mass fraction is 0.5% PluronicF-68, and mass fraction is 1% sorb
Alcohol, concentration are the MES buffer solutions of 0.1mol/L.
Preferably, the stabilizer of color developing agent includes tonyred 3 of the concentration for 35 μm of ol/L, and concentration is the Soviet Union of 20 μm of ol/L
Red red G, concentration are the carbohydrazide of 50 μm of ol/L, and concentration is the DTPA of 140 μm of ol/L, and mass fraction is 0.5% Pluronic
F-68, mass fraction are 1% sorbierite, and concentration is the MES buffer solutions of 0.1mol/L.
Preferably, the stabilizer provided in this embodiment is used for the stabilization of coloring agent TMB.
In this embodiment, the molar ratio of azo dyes and coloring agent TMB are 100:10000 or 55:10000.
In this embodiment, the suitable services pH value of the stabilizer of coloring agent is 6.0.
In one embodiment of the invention, the stabilizer of coloring agent include azo dyes, reducing agent, surfactant,
Auxiliary stabilizer and buffer；Wherein, the concentration of azo dyes is 45 μm of ol/L, and the concentration of reducing agent is 25 μm of ol/L, surface
The mass fraction of activating agent is 0.5%, and the mass fraction of auxiliary stabilizer is 1%, and the concentration of buffer is 0.1mol/L.
Preferably, azo dyes is acid red 18 and Eriochrome Red B, reducing agent is carbohydrazide；Surfactant is
Pluronic F-68；Auxiliary stabilizer is sorbierite；Buffer solution is MES buffer solutions.
Preferably, the stabilizer of color developing agent includes acid red 18 of the concentration for 40 μm of ol/L, and concentration is 5 μm of ol/L according to Lay
Chrome red B, concentration are the carbohydrazide of 25 μm of ol/L, and mass fraction is 0.5% Pluronic F-68, and mass fraction is 1% mountain
Pears alcohol, concentration are the MES buffer solutions of 0.1mol/L.
Preferably, the stabilizer provided in this embodiment is used for the stabilization of coloring agent TMB.
In this embodiment, the molar ratio of azo dyes and coloring agent TMB are 45:10000.
In this embodiment, the suitable services pH value of the stabilizer of coloring agent is 6.0.
In one embodiment of the invention, the stabilizer of coloring agent includes azo dyes, surfactant, auxiliary stabilization
Agent and buffer；Wherein, the concentration of azo dyes is 40 μm of ol/L, and the mass fraction of surfactant is 0.5%, and auxiliary is stablized
The mass fraction of agent is 1%, and the concentration of buffer is 0.1mol/L.
Preferably, azo dyes is acid red 18；Surfactant is Pluronic F-68；Auxiliary stabilizer is mountain
Pears alcohol；Buffer solution is MES buffer solutions.
Preferably, the stabilizer of color developing agent includes acid red 18 of the concentration for 40 μm of ol/L, and mass fraction is 0.5%
Pluronic F-68, mass fraction are 1% sorbierite, and concentration is the MES buffer solutions of 0.1mol/L.
Preferably, the stabilizer provided in this embodiment is used for the stabilization of coloring agent TMB.
In this embodiment, the molar ratio of azo dyes and coloring agent TMB are 40:10000.
In this embodiment, the suitable services pH value of the stabilizer of coloring agent is 6.0.
In one embodiment of the invention, the stabilizer of coloring agent includes azo dyes, reducing agent, chelating agent and buffering
Agent；Wherein, the concentration of azo dyes is 35 μm of ol/L~80 μm ol/L, and the concentration of reducing agent is 15 μm of ol/L~30 μm ol/L,
The concentration of chelating agent is 80 μm of ol/L~100 μm ol/L, and the concentration of buffer is 0.1mol/L.
Preferably, azo dyes is selected from lemon yellow, orange G or Eriochrome Red B；Reducing agent is paracetamol；Chela
Mixture is GEDTA；Buffer solution is phosphate buffer.
Preferably, the stabilizer of color developing agent includes lemon yellow of the concentration for 60 μm of ol/L；Concentration is orange for 20 μm of ol/L's
G；Concentration is the paracetamol of 30 μm of ol/L；Concentration is the GEDTA of 80 μm of ol/L；Concentration is that the phosphate of 0.1mol/L delays
Fliud flushing.
Preferably, the stabilizer of color developing agent includes Eriochrome Red B of the concentration for 15 μm of ol/L；Concentration is the orange of 20 μm of ol/L
Yellow G；Concentration is the paracetamol of 15 μm of ol/L；Concentration is the GEDTA of 100 μm of ol/L；Concentration is the phosphoric acid of 0.1mol/L
Salt buffer.
Preferably, the stabilizer provided in this embodiment is used for the stabilization of coloring agent TMB.
In this embodiment, the molar ratio of azo dyes and coloring agent TMB are 80:10000 or 35:10000.
In this embodiment, the suitable services pH value of the stabilizer of coloring agent is 6.0.
In one embodiment of the invention, the stabilizer of coloring agent includes azo dyes, reducing agent and buffer；Its
In, the concentration of azo dyes is 75 μm of ol/L, and the concentration of reducing agent is 20 μm of ol/L, and the concentration of buffer is 0.1mol/L.
Preferably, azo dyes is sudan red 7B and acid red 18；Reducing agent is paracetamol；Buffer solution is phosphorus
Phthalate buffer.
Preferably, the stabilizer of color developing agent includes sudan red 7B of the concentration for 35 μm of ol/L；Concentration is the acid of 40 μm of ol/L
Property red 18；Concentration is the paracetamol of 20 μm of ol/L；Concentration is the phosphate buffer of 0.1mol/L.
Preferably, the stabilizer provided in this embodiment is used for the stabilization of coloring agent TMB.
In this embodiment, the molar ratio of azo dyes and coloring agent TMB are 75:10000.
In this embodiment, the suitable services pH value of the stabilizer of coloring agent is 6.0.
In one embodiment of the invention, the stabilizer of coloring agent includes azo dyes, chelating agent and buffer；Its
In, the concentration of azo dyes is 40 μm of ol/L, and the concentration of chelating agent is 150 μm of ol/L, and the concentration of buffer is 0.1mol/L.
Preferably, azo dyes is acid red 18；Chelating agent is GEDTA；Buffer solution is phosphate buffer.
Preferably, the stabilizer of color developing agent includes acid red 18 of the concentration for 40 μm of ol/L；Concentration is 150 μm of ol/L
GEDTA；Concentration is the phosphate buffer of 0.1mol/L.
Preferably, the stabilizer provided in this embodiment is used for the stabilization of coloring agent TMB.
In this embodiment, the molar ratio of azo dyes and coloring agent TMB are 40:10000.
In this embodiment, the suitable services pH value of the stabilizer of coloring agent is 6.0.
In one embodiment of the invention, the stabilizer of coloring agent includes azo dyes, auxiliary stabilizer and buffer；
Wherein, the concentration of azo dyes is 55 μm of ol/L~115 μm ol/L, and the mass fraction of auxiliary stabilizer is 2%, buffer it is dense
Spend for 0.1mol/L.
Preferably, azo dyes is lemon yellow and orange G；Auxiliary stabilizer is BSA；Buffer solution is MES buffer solutions.
Preferably, the stabilizer of color developing agent includes lemon yellow of the concentration for 55 μm of ol/L；Mass fraction is 2% BSA；It is dense
Spend the MES buffer solutions for 0.1mol/L.
Preferably, the stabilizer of color developing agent includes lemon yellow of the concentration for 100 μm of ol/L；Concentration is orange for 15 μm of ol/L's
G；Mass fraction is 2% BSA；Concentration is the MES buffer solutions of 0.1mol/L.
Preferably, the stabilizer provided in this embodiment is used for the stabilization of coloring agent DHBS.
In this embodiment, the molar ratio of azo dyes and coloring agent DHBS are 55:10000 or 115:1000.
In this embodiment, the suitable services pH value of the stabilizer of coloring agent is 6.5.
In one embodiment of the invention, the stabilizer of coloring agent include azo dyes, reducing agent, auxiliary stabilizer and
Buffer；Wherein, the concentration of azo dyes is 100 μm of ol/L；The concentration of reducing agent is 50 μm of ol/L；The quality of auxiliary stabilizer
Fraction is 2%；The concentration of buffer is 0.1mol/L.
Preferably, azo dyes is direct red 2B；Auxiliary stabilizer is BSA；Buffer solution is MES buffer solutions.
Preferably, the stabilizer of color developing agent includes direct red 2B of the concentration for 100 μm of ol/L；Concentration is the 1- of 50 μm of ol/L
Carbaphen；Mass fraction is 2% BSA；Concentration is the MES buffer solutions of 0.1mol/L.
Preferably, the stabilizer provided in this embodiment is used for the stabilization of coloring agent DHBS.
In this embodiment, the molar ratio of azo dyes and coloring agent DHBS are 100:10000.
In this embodiment, the suitable services pH value of the stabilizer of coloring agent is 6.5.
In one embodiment of the invention, the stabilizer of coloring agent include azo dyes, chelating agent, auxiliary stabilizer and
Buffer；Wherein, the concentration of azo dyes is 150 μm of ol/L；The concentration of chelating agent is 20 μm of ol/L；The quality of auxiliary stabilizer
Fraction is 2%；The concentration of buffer is 0.1mol/L.
Preferably, azo dyes is tonyred 3；Chelating agent is EDTA；Auxiliary stabilizer is BSA；Buffer solution is MES
Buffer solution.
Preferably, the stabilizer of color developing agent includes tonyred 3 of the concentration for 150 μm of ol/L；Concentration is 20 μm of ol/L
EDTA；Mass fraction is 2% BSA；Concentration is the MES buffer solutions of 0.1mol/L.
Preferably, the stabilizer provided in this embodiment is used for the stabilization of coloring agent DHBS.
In this embodiment, the molar ratio of azo dyes and coloring agent DHBS are 150:10000.
In this embodiment, the suitable services pH value of the stabilizer of coloring agent is 6.5.
In one embodiment of the invention, the stabilizer of coloring agent include azo dyes, reducing agent, surfactant,
Auxiliary stabilizer and buffer；Wherein, the concentration of azo dyes is 10 μm of ol/L；The concentration of reducing agent is 50 μm of ol/L；Surface
The mass fraction of activating agent is 0.5%；The mass fraction of auxiliary stabilizer is 2%；The concentration of buffer is 0.1mol/L.
Preferably, azo dyes is acid red 66；Reducing agent is 4- Carbaphens；Surfactant is Twenn
20；Auxiliary stabilizer is trehalose；Buffer solution is MOPS buffer solutions.
Preferably, the stabilizer of color developing agent includes acid red 66 of the concentration for 10 μm of ol/L；Concentration is the 4- of 50 μm of ol/L
Carbaphen；Mass fraction is 0.5% Twenn 20；Mass fraction is 2% trehalose；Concentration is 0.1mol/L's
MOPS buffer solutions.
Preferably, the stabilizer provided in this embodiment is used for the stabilization of coloring agent TBHBA.
In this embodiment, the molar ratio of azo dyes and coloring agent TBHBA are 10:10000.
In this embodiment, the suitable services pH value of the stabilizer of coloring agent is 7.4.
In one embodiment of the invention, the stabilizer of coloring agent includes azo dyes, surfactant, auxiliary stabilization
Agent and buffer；Wherein, the concentration of azo dyes is 125 μm of ol/L；The mass fraction of surfactant is 0.5%；Auxiliary is steady
The mass fraction for determining agent is 2%；The concentration of buffer is 0.1mol/L.
Preferably, azo dyes is direct red 2B and lemon yellow；Surfactant is Twenn 20；Auxiliary stabilizer is
Trehalose；Buffer solution is MOPS buffer solutions.
Preferably, the stabilizer of color developing agent includes direct red 2B of the concentration for 25 μm of ol/L；Concentration is the lemon of 100 μm of ol/L
Lemon is yellow；Mass fraction is 0.5% Twenn 20；Mass fraction is 2% trehalose；The MOPS that concentration is 0.1mol/L is buffered
Liquid.
Preferably, the stabilizer provided in this embodiment is used for the stabilization of coloring agent TBHBA.
In this embodiment, the molar ratio of azo dyes and coloring agent TBHBA are 125:10000.
In this embodiment, the suitable services pH value of the stabilizer of coloring agent is 7.4.
In one embodiment of the invention, the stabilizer of coloring agent includes azo dyes, surfactant, auxiliary stabilization
Agent and buffer；Wherein, the concentration of azo dyes is 85 μm of ol/L；The mass fraction of surfactant is 0.5%；Auxiliary is stablized
The mass fraction of agent is 2%；The concentration of buffer is 0.1mol/L.
Preferably, azo dyes is sunset yellow；Surfactant is Twenn 20；Auxiliary stabilizer is trehalose；It is slow
Fliud flushing is MOPS buffer solutions.
Preferably, the stabilizer of color developing agent includes sunset yellow of the concentration for 85 μm of ol/L；Mass fraction is 0.5%
Twenn 20；Mass fraction is 2% trehalose；Concentration is the MOPS buffer solutions of 0.1mol/L.
Preferably, the stabilizer provided in this embodiment is used for the stabilization of coloring agent TBHBA.
In this embodiment, the molar ratio of azo dyes and coloring agent TBHBA are 85:10000.
In this embodiment, the suitable services pH value of the stabilizer of coloring agent is 7.4.
In an embodiment of the present invention, the mass ratio of color developing agent and the stabilizer of color developing agent provided by the invention is 1:(10
~100000).
In certain embodiments, the mass ratio of color developing agent and the stabilizer of color developing agent provided by the invention is 1:(100~
10000)。
Application of the stabilizer of color developing agent provided by the invention in biochemistry detection reagent is prepared；Biochemistry detection is red to courage
Element, hemoglobin, glucose, creatinine, uric acid, cholesterol, triglyceride, high-density lipoprotein cholesterol, glutamic-pyruvic transaminase or
The quantitative or qualitative detection of glutamic-oxalacetic transaminease.
Need to add corresponding oxidizing ferment when measuring above-mentioned analyte, to produce hydrogen peroxide and above-mentioned chromogenic reagent.
For example, add glucose oxidase during measure glucose.
The stabilizer of the present invention can also be applied to dry chemical and detect.The composition of the present invention is fixed on solid phase carrier
On, then dry.Sample is added on solid phase carrier during test, is reacted.Good stability can equally be obtained.
The present invention uses stabilizer of the azo dyes as color developing agent, has not only acted as good stabilization, and not
Detection can be interfered.Experiment shows, states the addition of azo dyes, improves the spontaneous colour developing of color developing agent and makes color developing agent
Maintain more preferable coloration ability.In the check experiment of azo dyes is not added, all there occurs a degree of face for coloring agent
Color changes, and also there occurs obvious rise, and after using stabilizer provided by the invention, dye color changes unknown light absorption value
Aobvious, light absorption value change is not notable.Also, after high temperature is placed for a long time, remain to obtain good detection result.
Embodiment
Stabilizer and its application the present invention provides a kind of color developing agent, those skilled in the art can be used for reference in this paper
Hold, be suitably modified technological parameter realization.In particular, all similar substitutions and modifications are to those skilled in the art
For be it will be apparent that they are considered as being included in the present invention.The method of the present invention and application are by preferably implementing
Example is described, related personnel substantially can not depart from present invention, in spirit and scope to methods herein and application
It is modified or suitably changes with combining, realizes and using the technology of the present invention.
The reagent or instrument that the present invention uses are all common commercially available product, can all be bought in market.
With reference to embodiment, the present invention is further explained：
Embodiment 1
It is formulated as follows the composition shown in table：
1 embodiment of table, 1 experimental program
| Option A | Option b | Scheme C | Scheme D | Scheme E | |
| TMB | 10mmol/L | 10mmol/L | 10mmol/L | 10mmol/L | 10mmol/L |
| Sudan orange B | 1μmol/L | ||||
| Alizarin yellow R | 10μmol/L | ||||
| Orange G | 5μmol/L | ||||
| Lemon yellow | 5μmol/L | ||||
| Luo Yellow | 20μmol/L | ||||
| Crocien orange G | 100μmol/L | ||||
| TritonX100 | 1% | 1% | 1% | 1% | 1% |
| MOPS | 0.1mol/L | 0.1mol/L | 0.1mol/L | 0.1mol/L | 0.1mol/L |
PH value is adjusted to 7.4 with hydrochloric acid or sodium hydroxide.
The absorbance of 650nm is 0.02 after each solution is prepared.50 DEG C are placed 7 days, test the extinction of the 650nm of above-mentioned solution
Degree, it is as a result as shown in the table：
Absorbance after 2 embodiment of table, 1 each 50 DEG C of scheme is placed 7 days
| Option A | Option b | Scheme C | Scheme D | Scheme E | |
| Absorbance | 0.201 | 0.156 | 0.143 | 0.135 | 0.137 |
In addition, taking above-mentioned solution 1ml, the 100 μ l of HRP solution of 20U/L are added, add the 100 μ l of hydrogen peroxide of 10mmol/L,
37 DEG C of hatching 3min, observe the color of above-mentioned solution, as a result as shown in the table：
Color after 3 embodiment of table, 1 each 50 DEG C of scheme is placed 7 days
| Option A | Option b | Scheme C | Scheme D | Scheme E | |
| Color | Pale blue | It is blue | It is blue | It is blue | It is blue |
As it can be seen that the addition of above-mentioned dyestuff, improves the spontaneous colour developing of TMB and color developing agent is maintained preferably colour developing energy
Power.
Embodiment 2
It is formulated as follows the composition shown in table：
4 embodiment of table, 2 experimental program
| Option A | Option b | Scheme C | Scheme D | Scheme E | |
| 4AAP | 10mmol/L | 10mmol/L | 10mmol/L | 10mmol/L | 10mmol/L |
| MAOS | 10mmol/L | 10mmol/L | 10mmol/L | 10mmol/L | 10mmol/L |
| Acid red 66 | -- | 0.1μmol/L | |||
| Sudan black B | -- | 50μmol/L | |||
| Alkaline brown g | -- | 15μmol/L | |||
| Tonyred B | -- | 75μmol/L | |||
| Direct black BN | -- | 40μmol/L | |||
| Sudan red 7B | -- | 1mmol/L | |||
| Phosphate buffer | 0.1mol/L | 0.1mol/L | 0.1mol/L | 0.1mol/L | 0.1mol/L |
PH value is adjusted to 6.5 with hydrochloric acid or sodium hydroxide.
The absorbance of 630nm is 0.015 after each solution is prepared.60 DEG C are placed 7 days, test the suction of the 630nm of above-mentioned solution
Luminosity, it is as a result as shown in the table：
Absorbance after 5 embodiment of table, 2 each 60 DEG C of scheme is placed 7 days
| Option A | Option b | Scheme C | Scheme D | Scheme E | |
| Absorbance | 0.101 | 0.045 | 0.063 | 0.071 | 0.083 |
In addition, taking above-mentioned solution 1ml, the 100 μ l of HRP solution of 20U/L are added, add the 100 μ l of hydrogen peroxide of 10mmol/L,
37 DEG C of hatching 3min, observe the color of above-mentioned solution, as a result as shown in the table：
Color after 6 embodiment of table, 2 each 60 DEG C of scheme is placed 7 days
| Option A | Option b | Scheme C | Scheme D | Scheme E | |
| Color | Pale blue | It is blue | It is blue | It is blue | It is blue |
As it can be seen that the addition of above-mentioned dyestuff, improves the spontaneous colour developing of 4AAP and MAOS and color developing agent is maintained more preferably
Coloration ability.
Embodiment 3
It is formulated as follows the composition shown in table：
7 embodiment of table, 3 experimental program
| Option A | Option b | Scheme C | Scheme D | Scheme E | |
| 4AAP | 10mmol/L | 10mmol/L | 10mmol/L | 10mmol/L | 10mmol/L |
| MADB | 10mmol/L | 10mmol/L | 10mmol/L | 10mmol/L | 10mmol/L |
| Direct red 2B | -- | 10μmol/L | |||
| Tonyred 3 | -- | 20μmol/L | |||
| Tonyred 4 | -- | 35μmol/L | |||
| Lure red | -- | 15μmol/L | |||
| EDTA | -- | 40μmol/L | |||
| 4- Carbaphens | -- | 50μmol/L | 100μmol/L | ||
| Sucrose | 2% | 2% | 2% | 2% | 2% |
| HEPES | 0.1mol/L | 0.1mol/L | 0.1mol/L | 0.1mol/L | 0.1mol/L |
PH value is adjusted to 7.5 with hydrochloric acid or sodium hydroxide.
The absorbance of 630nm is 0.014 after each solution is prepared.60 DEG C are placed 7 days, test the suction of the 630nm of above-mentioned solution
Luminosity, it is as a result as shown in the table：
Absorbance after 8 embodiment of table, 3 each 60 DEG C of scheme is placed 7 days
| Option A | Option b | Scheme C | Scheme D | Scheme E | |
| Absorbance | 0.125 | 0.035 | 0.045 | 0.077 | 0.047 |
In addition, taking above-mentioned solution 1ml, the 100 μ l of HRP solution of 20U/L are added, add the 100 μ l of hydrogen peroxide of 10mmol/L,
37 DEG C of hatching 3min, observe the color of above-mentioned solution, as a result as shown in the table：
Color after 9 embodiment of table, 3 each 60 DEG C of scheme is placed 7 days
| Option A | Option b | Scheme C | Scheme D | Scheme E | |
| Color | Pale blue | It is blue | It is blue | It is blue | It is blue |
As it can be seen that the addition of above-mentioned dyestuff, improves the spontaneous colour developing of 4AAP and MADB and color developing agent is maintained more preferably
Coloration ability.
Embodiment 4
It is formulated as follows the composition shown in table：
10 embodiment of table, 4 experimental program
PH value is adjusted to 6.0 with hydrochloric acid or sodium hydroxide.
The absorbance of 650nm is 0.018 after each solution is prepared.70 DEG C are placed 7 days, test the suction of the 650nm of above-mentioned solution
Luminosity, it is as a result as shown in the table：
Absorbance after 11 embodiment of table, 4 each 70 DEG C of scheme is placed 7 days
| Option A | Option b | Scheme C | Scheme D | Scheme E | |
| Absorbance | 0.205 | 0.103 | 0.095 | 0.085 | 0.071 |
In addition, taking above-mentioned solution 1ml, the 100 μ l of HRP solution of 20U/L are added, add the 100 μ l of hydrogen peroxide of 10mmol/L,
37 DEG C of hatching 3min, observe the color of above-mentioned solution, as a result as shown in the table：
Color after 12 embodiment of table, 4 each 70 DEG C of scheme is placed 7 days
| Option A | Option b | Scheme C | Scheme D | Scheme E | |
| Color | Pale blue | It is blue | It is blue | It is blue | It is blue |
As it can be seen that the addition of above-mentioned dyestuff, improves the spontaneous colour developing of TMB and color developing agent is maintained preferably colour developing energy
Power.
Embodiment 5
It is formulated as follows the composition shown in table：
13 embodiment of table, 5 experimental program
| Option A | Option b | Scheme C | Scheme D | Scheme E | |
| TMB | 10mmol/L | 10mmol/L | 10mmol/L | 10mmol/L | 10mmol/L |
| Lemon yellow | 60μmol/L | ||||
| Sudan red 7B | 35μmol/L | ||||
| Acid red 18 | 40μmol/L | 40μmol/L | |||
| Eriochrome Red B | 15μmol/L | ||||
| Orange G | 20μmol/L | 20μmol/L | |||
| GEDTA | 80μmol/L | 100μmol/L | 150μmol/L | ||
| Paracetamol | 30μmol/L | 20μmol/L | 15μmol/L | ||
| Phosphate buffer | 0.1mol/L | 0.1mol/L | 0.1mol/L | 0.1mol/L | 0.1mol/L |
PH value is adjusted to 6.0 with hydrochloric acid or sodium hydroxide.
The absorbance of 650nm is 0.019 after each solution is prepared.60 DEG C are placed 7 days, test the suction of the 650nm of above-mentioned solution
Luminosity, it is as a result as shown in the table：
Absorbance after 14 embodiment of table, 5 each 60 DEG C of scheme is placed 7 days
| Option A | Option b | Scheme C | Scheme D | Scheme E | |
| Absorbance | 0.205 | 0.030 | 0.045 | 0.025 | 0.021 |
In addition, taking above-mentioned solution 1ml, the 100 μ l of HRP solution of 20U/L are added, add the 100 μ l of hydrogen peroxide of 10mmol/L,
37 DEG C of hatching 3min, observe the color of above-mentioned solution, as a result as shown in the table：
Color after 15 embodiment of table, 5 each 60 DEG C of scheme is placed 7 days
| Option A | Option b | Scheme C | Scheme D | Scheme E | |
| Color | Pale blue | It is blue | It is blue | It is blue | It is blue |
As it can be seen that the addition of above-mentioned dyestuff, improves the spontaneous colour developing of TMB and color developing agent is maintained preferably colour developing energy
Power.
Embodiment 6
It is formulated as follows the composition shown in table：
16 embodiment of table, 6 experimental program
PH value is adjusted to 6.5 with hydrochloric acid or sodium hydroxide.
Above-mentioned 5 schemes are each in 2-8 DEG C and 45 DEG C of temperature respectively to place portion, respectively with 2-8 in 5 schemes after 3 days
DEG C preserve solution testing 2.5mmol/L, 8.7mmol/L, 11mmol/L T-CHOL serum sample (above-mentioned total cholesterol concentration
Measured using full automatic biochemical apparatus), obtain absorbance.Standard curve is drawn according to concentration of specimens and obtains regression equation.
Test method is to add 0.3ml serum samples in the above-mentioned solution of 2.7ml, and 37 DEG C of hatching 5min, read in 520nm and inhale
Luminosity.
Above-mentioned 5 schemes are tested into above-mentioned 3 samples after the same method in 60 DEG C of storage solutions, by absorbance result
Bring above-mentioned regression equation into, test sample concentration is calculated.As a result (unit as shown in the table：mmol/L)：
17 embodiment of table, 6 each scheme absorbance
| Sample actual concentrations | Option A | Option b | Scheme C | Scheme D | Scheme E |
| 2.5mmol/L | 3.5 | 2.8 | 2.7 | 2.9 | 2.7 |
| 8.7mmol/L | 6.3 | 8.2 | 8.2 | 7.8 | 7.9 |
| 11mmol/L | 7.5 | 10.5 | 3.7 | 10.1 | 10.2 |
As can be seen from the above table, the protectant option A of the present invention is not added, and low value (2.5mmol/L) is higher, is because the back of the body
Scape rise causes.And it is due to that color developing agent goes bad and loses coloration ability that high level (11mmol/L), which reduces,.And add the present invention's
After protective agent, the problem of these two aspects, is obtained for significant improvement, that is, improves the stability of color developing agent.
Embodiment 7
It is formulated as follows the composition shown in table：
18 embodiment of table, 7 experimental program
PH value is adjusted to 7.4 with hydrochloric acid or sodium hydroxide.
Above-mentioned 5 schemes are coated on the filter paper of 1cm × 1cm, each scheme applies 2 filter paper.2h is toasted at 40 DEG C.Drying
Two filter paper are respectively put into 2-8 DEG C and 55 DEG C of temperature afterwards, after 7 days.Respectively with the filter paper of 2-8 DEG C of preservation in 5 schemes
Testing 0.11mmol/L, 0.67mmol/L, 1.1mmol/L uric acid serum sample, (above-mentioned uric acid concentration uses full automatic biochemical apparatus
Measure), obtain reflectivity.Standard curve is drawn according to concentration of specimens and obtains regression equation.
For 30 μ l serum samples are added dropwise on filter paper, 37 DEG C of hatching 3min, read 530nm and read reflectivity test method.
Above-mentioned 5 schemes are stored into filter paper at 60 DEG C and test above-mentioned 3 samples after the same method, by reflectivity results
Bring above-mentioned regression equation into, test sample concentration is calculated.As a result (unit as shown in the table：mmol/L)：
19 embodiment of table, 7 each scheme measured concentration
| Sample actual concentrations | Option A | Option b | Scheme C | Scheme D | Scheme E |
| 0.11mmol/L | 0.20 | 0.13 | 0.14 | 0.13 | 0.12 |
| 0.67mmol/L | 0.55 | 0.65 | 0.66 | 0.62 | 0.68 |
| 1.1mmol/L | 0.75 | 1.02 | 1.05 | 1.03 | 1.12 |
As can be seen from the above table, do not add the protectant option A of the present invention, low value (0.11mmol/L) is higher, be because
Background rise causes.And it is due to that color developing agent goes bad and loses coloration ability that high level (1.1mmol/L), which reduces,.And add the present invention
Protective agent after, the problem of these two aspects, is obtained for significant improvement, that is, improves the stability of color developing agent.
It the above is only the preferred embodiment of the present invention, it is noted that come for those skilled in the art
Say, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications also should be regarded as
Protection scope of the present invention.
Claims (8)
1. a kind of composition, it is characterised in that including color developing agent and stabilizer：
The stabilizer includes azo dyes, further includes in reducing agent, chelating agent, buffer, auxiliary stabilizer, surfactant
It is any or both more than combination；
The color developing agent is at least one of TMB, 4AAP, MAOS, MADB, DHBS, TBHBA；
The azo dyes is selected from monoazo dyes, bisazo dye or polyazo dye；
The monoazo dyes be selected from tonyred l, tonyred 2, sudan orange B, alizarin yellow R, orange G, tonyred G, according to Lay
Chrome red B, acid red 18 or crocien orange G；
The bisazo dye is selected from acid red 66, Sudan black B, directly red 2B, alkaline brown g, tonyred B, tonyred 3, Soviet Union
Red red No. 4 or sudan red 7B；
The polyazo dye is direct black BN；
The reducing agent be selected from 1- Carbaphens, 1,5- diphenyl ureas, N, N, N- trimethylamino ureas, 4- Carbaphens,
Bisphenol-A, gallic acid, carbohydrazide, paracetamol or gentianic acid；
The chelating agent be selected from ethylenediamine tetra-acetic acid, diethylene-triamine pentaacetic acid, anti-form-1,2- diaminocyclohexane-N, N,
N ', N '-tetraacethyl, O, O '-two (2- aminoethyls) ethylene glycol-N, N, N ', N '-tetraacethyl or nitrilotriacetic acid；
The buffer is selected from citric acid buffer agent, phosphate buffer, acetate buffer agent or Good ' s buffers；
The surfactant is selected from Tween series, Triton series, Brij series or Pluronic series；
The auxiliary stabilizer is selected from carbohydrate or protide.
2. composition according to claim 1, it is characterised in that the concentration of wherein described reducing agent for 10 μm of ol/L~
100μmol/L。
3. composition according to claim 1, it is characterised in that the concentration of wherein described chelating agent for 20 μm of ol/L~
150μmol/L。
4. composition according to claim 1, it is characterised in that the concentration of wherein described buffer is 0.1mol/L.
5. composition according to claim 1, it is characterised in that the mass fraction of wherein described surfactant is
0.5%~1%.
6. composition according to claim 1, it is characterised in that the mass fraction of wherein described auxiliary stabilizer is 1%
~2%.
7. according to claim 1~6 any one of them composition, it is characterised in that the color developing agent and claim 1~6
The mass ratio of the stabilizer of any one of them color developing agent is 1:(10~100000).
8. application of claim 1~6 any one of them composition in biochemistry detection reagent is prepared；The biochemistry detection is
To the quantitative or qualitative detection of glucose, creatinine, uric acid, cholesterol, triglyceride, glutamic-pyruvic transaminase or glutamic-oxalacetic transaminease.
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| CN106771130B (en) * | 2016-11-17 | 2018-12-28 | 陕西师范大学 | A kind of enzyme-linked immune analytic method |
| CN108918415B (en) * | 2018-07-19 | 2021-09-03 | 美康生物科技股份有限公司 | Human total oxidation resistance state detection kit |
| CN110791122A (en) * | 2019-11-28 | 2020-02-14 | 迈克生物股份有限公司 | Sudan black B staining kit |
| CN112029817B (en) * | 2020-07-13 | 2024-02-02 | 兰州百源基因技术有限公司 | Creatinine detection kit and application method thereof |
| CN114184452B (en) * | 2021-11-30 | 2024-03-01 | 阿里生物技术泰州有限公司 | Dye bead for centrifugal microfluidic disc chip and preparation method thereof |
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