CN106771132A - The detection kit of sulfamethazine in a kind of food - Google Patents
The detection kit of sulfamethazine in a kind of food Download PDFInfo
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- CN106771132A CN106771132A CN201611027949.6A CN201611027949A CN106771132A CN 106771132 A CN106771132 A CN 106771132A CN 201611027949 A CN201611027949 A CN 201611027949A CN 106771132 A CN106771132 A CN 106771132A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/531—Production of immunochemical test materials
- G01N33/532—Production of labelled immunochemicals
- G01N33/535—Production of labelled immunochemicals with enzyme label or co-enzymes, co-factors, enzyme inhibitors or enzyme substrates
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
- G01N33/54326—Magnetic particles
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- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
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Abstract
The invention discloses a kind of detection kit of sulfamethazine in food, including：Liquid, concentrated cleaning solution are redissolved in enzyme-labelled antigen, enzyme-labelled antigen dilution, magnetic labeling antibody, magnetic labeling antibody dilution, sulfamethazine serial standards solution, chemical luminous substrate A liquid, chemical luminous substrate B liquid, concentration.Kit of the present invention has sensitivity and specificity higher, and the detection sensitivity to sulfamethazine can reach 0.025 μ g/L.
Description
Technical field
The present invention relates to technical field of food detection, the detection reagent of sulfamethazine in specifically a kind of food
Box.
Background technology
Sulfamethazine（Sulfamethazine, SMZ）Be broad spectrum antimicrobicide, to most of gram-positive bacterias and
Gram-negative bacteria is all inhibited, is clinically mainly used in treating various hemolytic streptococcus, meningococcus, pneumonia
Coccus etc. infects the bacteriosises such as triggered respiratory tract infection, enteric infection, thus is widely used in livestock-raising,
Residual phenomena in animal food is more serious.Acted in vivo due to sulfamethazine and the metabolism time is more long, led to
The sulfamethazine for crossing any approach intake is likely to be accumulated in human body, during more than finite concentration, will be to human body
Cause damage, it is long-term or be excessively used the damage that can cause urinary system, hemopoietic system, nervous system etc., or even have carcinogenic
Possibility.The country such as China, European Union and mechanism specify that sulfa drugs total amount and sulfanilamide (SN) dimethyl are phonetic in animal food
The MRL of the single medicine such as pyridine is 100 μ g/kg.
At present, the detection method for sulfamethazine residual has thin-layer chromatography, high performance liquid chromatography, liquid matter to join
With, gas-chromatography, gas chromatography mass spectrometry, Enzyme-linked Immunosorbent Assay（ELISA）, colloid gold immune technology etc..These methods all respectively have excellent lacking
Point, traditional liquid phase chromatography analytical method can only once detect a sample, because its pretreatment process is complicated, instrumentation degree
It is high, cumbersome, so general only make confirmatory test in the more testing agencies of funds.And common government's quality inspection organization,
Masses' laboratories such as enterprise, using it is most be ELISA method.ELISA method is by multiple elution process, label enzyme
Easy in inactivation, substrate is shown in that light is easily decomposed, the shortcomings of environmental disturbances factor is complicated.Colloidal gold chromatography can meet scene and quickly
Ask, but can only qualitative detection cannot obtain quantitative result.
The content of the invention
It is phonetic it is an object of the invention to provide sulfanilamide (SN) dimethyl in a kind of food with sensitivity and specificity higher
The detection kit of pyridine.
To achieve the above object, the present invention provides following technical scheme：
The detection kit of sulfamethazine in a kind of food, including：Enzyme-labelled antigen, enzyme-labelled antigen dilution, magnetic mark resist
Body, magnetic labeling antibody dilution, sulfamethazine serial standards solution, chemical luminous substrate A liquid, chemical luminous substrate B
Liquid, concentrated cleaning solution are redissolved in liquid, concentration.
As further scheme of the invention：Described enzyme-labelled antigen is the sulfanilamide (SN) dimethyl of horseradish peroxidase-labeled
The label of pyrimidine haptens, described sulfamethazine haptens is in methyl alcohol by sulfamethazine and ethylenediamine
Reacted in solution and obtained.
As further scheme of the invention：Described enzyme-labelled antigen be stored in the tween containing 0.055~0.06wt.%-
20th, in the phosphate buffer of the pH value 7.2~7.4 of 0.025~0.028mol/L.
As further scheme of the invention：Described enzyme-labelled antigen dilution is Na3PO4Concentration be 0.014mol/L,
NaCl concentration is the cushioning liquid of the pH value 7.2~7.4 of 0.20mol/L.
As further scheme of the invention：Described magnetic labeling antibody is sulfamethazine monoclonal antibody and silicon substrate
Magnetic bead coupling is obtained；Content 0.l~the 0.3eq/g of described silicon substrate magnetic bead surfaces group, described magnetic labeling antibody is stored in and contains
In the Tween-20 of 0.35~0.4wt.%, the phosphate buffer of the pH value 7.2~7.4 of 0.035~0.04mol/L.
As further scheme of the invention：Described magnetic labeling antibody dilution is Na2HPO4Concentration be 0.012mol/L,
NaCl concentration is the cushioning liquid of the pH value 7.2~7.4 of 0.25mol/L.
As further scheme of the invention：Described sulfamethazine monoclonal antibody is phonetic by sulfanilamide (SN) dimethyl
The conjugate that pyridine haptens is obtained with chicken ovalbumin is prepared as immunogen immune Balb/c mouse.
As further scheme of the invention：Described sulfamethazine monoclonal antibody is phonetic by sulfanilamide (SN) dimethyl
Pyridine haptens is thin as immunogen immune Balb/c mouse, cell fusion, hybridoma with the conjugate that chicken ovalbumin is obtained
The screening of born of the same parents, the titration of subclone and mouse ascites are obtained.
As further scheme of the invention：Described chemical luminous substrate A liquid be containing luminol, p-cresol three
Hydroxymethyl aminomethane solution, described chemical luminous substrate B liquid is to contain sodium citrate, anhydrous Na2HPO4With CO (NH2)2·
H2O2The aqueous solution.
As further scheme of the invention：Described chemical luminous substrate A liquid be luminol content be 0.026~
0.028 μ g/L, p-cresol content are 0.04~0.0042 μ g/L, the tris solution of pH value 8.4~8.6,
Described chemical luminous substrate B liquid is that every 100ml aqueous solution contains 2~2.2g of sodium citrate, anhydrous Na2HPO43~3.5g and body
Product percentage composition is 0.65% CO (NH2)2·H2O21.5~1.8ml.
As further scheme of the invention：Described sulfamethazine serial standards solution concentration is respectively：0
μ g/L, 0.025 μ g/L, 0.05 μ g/L, 0.1 μ g/L, 0.2 μ g/L, 0.4 μ g/L, 0.8 μ g/L, 1.6 μ g/L, standard dilutions
It is Tween-20 containing 0.035wt.%, the phosphate buffer of the pH value 7.4 of 0.042mol/L.
As further scheme of the invention：It is that liquid is redissolved in 20 times of concentrations that liquid is redissolved in described concentration, and specifically every liter contains
The NaH of 45~60g2PO4·2H2O, 200~220g Na2HPO4·12H2The aqueous solution of O.
As further scheme of the invention：Described concentrated cleaning solution is 20 times of concentrated cleaning solutions, specifically containing 0.38
The phosphate buffer of~0.42wt.% Tween-20s, the pH value 7.2~7.4 of 2.6~3mol/L.
The method detected using the detection kit of sulfamethazine in described food, including following step
Suddenly：
（1）By enzyme-labelled antigen and enzyme-labelled antigen dilution according to 1：20 volume ratio is diluted, and obtains enzyme-labelled antigen working solution；
（2）By magnetic labeling antibody and magnetic labeling antibody dilution according to 1：20 volume ratio is diluted, and obtains magnetic labeling antibody working solution；
（3）The work of 30~35 μ L enzyme-labelled antigens working solutions, 30~35 μ L samples extract solutions and 30~35 μ L magnetic labeling antibodies is taken respectively
Liquid, is added sequentially in container, and 18min is reacted at room temperature, then carries out 2~3min of Magneto separate, after abandoning supernatant, uses cleaning solution
500~800 μ L are cleaned 3~4 times to complex precipitate；
（4）Chemical luminous substrate A liquid and each 30 μ L of chemical luminous substrate B liquid are added toward the compound of separator well, detection sends
Relative light intensity（RLU）, the content of sulfamethazine and RLU, can be by RLU standards into negative correlativing relation in sample
Curve calculates the residual concentration of sulfamethazine.
Compared with prior art, the beneficial effects of the invention are as follows：Kit of the present invention is with sensitivity higher and specifically
Property, the detection sensitivity to sulfamethazine can reach 0.025 μ g/L.
Specific embodiment
Below in conjunction with the embodiment of the present invention, the technical scheme in the embodiment of the present invention is clearly and completely described,
Obviously, described embodiment is only a part of embodiment of the invention, rather than whole embodiments.Based in the present invention
Embodiment, the every other embodiment that those of ordinary skill in the art are obtained under the premise of creative work is not made, all
Belong to the scope of protection of the invention.
Embodiment 1：The preparation of kit concrete component
1st, sulfamethazine hapten synthesis
0.58g sulfamethazines and mixed liquor of the 0.082g ethylenediamines in 80ml methyl alcohol, react 4~5h at room temperature,
Solvent is evaporated off, sulfamethazine haptens is quantitatively obtained.
2nd, the preparation of enzyme-labelled antigen
10~15mg sulfamethazine haptens is taken, 1~1.5ml DMFs are dissolved in（DMF）In；Take
30~35mg dichloroethanes（EDC）And N-hydroxy-succinamide（NHS）Fully dissolved after addition half with 0.2~0.3ml water
In antigen lysate, 20h is stirred at room temperature, you can obtain reaction solution A；Weigh horseradish peroxidase（HRP）35~45mg, makes
Be substantially dissolved in the phosphate buffer of pH value 7.2,4ml, reaction solution A is dropwise slowly dropped in HRP solution, and in
20h is stirred at room temperature；Dialysed 3 days in 4 DEG C with the phosphate buffer of 0.015mol/L, 3 dialyzates are changed daily, to remove not
The small-molecule substance of reaction, obtains sulfamethazine enzyme-labelled antigen；Packing, saves backup in -20 DEG C.
3rd, the preparation of immunogene
35~45mg of HRP are replaced with into chicken ovalbumin（OVA）50~55mg, preparation method ibid, obtains immunogene.
4th, the preparation of sulfamethazine monoclonal antibody
A）Animal immune：It is complete with physiological saline solution immunogene and Freund with the above-mentioned immunogene prepared by 100 μ g/
Adjuvant is mixed in equal volume, and nape part hypodermic injection is immunized 6~8 week old Balb/c raettins, after initial immunity the 7th, 14,28 days in order to avoid
Epidemic focus is mixed in equal volume with incomplete Freund's adjuvant, and each supplementary immunization once, merges first 3 days with the μ g/ of immune complex 100,
Supplementary immunization is once again to be not added with Freund's adjuvant.
B）Cell fusion：Carry out according to a conventional method, take the splenocyte and the Mouse Bone in exponential phase of immune mouse
Myeloma cells（SP2/0）Mixing, was then slowly added to the fusion agent of preheating in 45 seconds（Polyethylene glycol 16000）Merged,
Suspended with HAT culture mediums uniform, add appropriate feeder cells, 96 well culture plates are incubated at, in 37 DEG C, 5%CO2Incubator
Middle culture, liquid is partly changed after 5 days with HT culture mediums, and liquid is changed full at 9 days.
C）The screening of hybridoma：After cell fusion, when cell grows to the 1/4 of culture hole area, sieved using substep
Method is selected to screen hybridoma.Primary election uses indirect ELISA method, with envelope antigen（In advance with square formation method conventional titration its most
Good coating concentration and positive serum dilution factor）Coated elisa plate, adds measured hole culture supernatant, is incubated, and sulfanilamide (SN) is added after cleaning
The μ L of dimethyl pyrimidine serial standards solution 30, add the μ L of the cell supernatant 30 and μ L of sheep anti-mouse igg-HRP 30, in 37 DEG C
Reaction 40min, board-washing adds the μ L of substrate solution nitrite ion 80, in lucifuge reaction 20min at 25 DEG C, adds the μ L of terminate liquid 20,
Determine OD450nmValue drops to less than the 50% of control wells, is judged to the positive, is all positive hole through 2~3 detections, immediately with limited
Dilution method carries out subcloning.
D）It is prepared by monoclonal antibody：2~3 times are subcloned the hybridoma Amplification Culture built after strain, collect supernatant
Potency is determined with indirect ELISA, is frozen；And take 8~10 week old Balb/c mouse peritoneal injection atoleines 0.5ml/ only, 7~
Intraperitoneal injection hybridoma 1~2 × 10 after 10 days6/ only, and mouse ascites are extracted after 7~10 days, centrifuging and taking supernatant determines effect
Valency, and freeze standby.
5th, the preparation of magnetic labeling antibody
A）Magnetic bead is activated
Silicon substrate magnetic bead, its activity group content is 0.1~0.3eq/g, takes 80 μ L silicon substrate magnetic beads, is distributed in glycerine at ultrasound
After reason, sodium citrate is added, after stirring, succinic anhydride is added dropwise, 12~16h is reacted after completion of dropping；After reaction terminates, Magneto separate
Go out carboxyl magnetic bead, be for several times successively 6~7 to cleaning solution pH value with absolute ethyl alcohol and deionized water cyclic washing sediment, ultrasound
20~25min for the treatment of, is dispersed in 2/10000ths NaN3In water, that is, obtain carboxyl magnetic bead；Carboxyl magnetic bead MES buffer solution for cleaning is extremely
Less once, in MES buffer solutions, after by activator carbodiimide, magnetic bead is activated, obtain to surface has carboxylic to resuspended carboxyl magnetic bead
The magnetic bead of base activation.
B）Magnetic bead is coupled the preparation of sulfamethazine monoclonal antibody
10~12 μ g sulfamethazine monoclonal antibodies are dissolved into the MES of the pH value 5.6,20mmol/L of 50 μ L, to
The magnetic bead of 3~4mg activation is wherein added, and cumulative volume is adjusted to 100 μ L with above-mentioned concentration MES solution, gently mix magnetic bead
With sulfamethazine monoclonal antibody；35~40min or 4 DEG C of coupling 2h is coupled under room temperature condition, whirlpool can be utilized during this period
Rotation instrument makes magnetic bead keep mixing state；Centrifuge tube is placed in and 3~4min of Magneto separate is carried out on Magneto separate frame, removes supernatant；In order to
Unreacted-COOH is quenched, the trishydroxymethylaminomethane of the pH value 7.2~7.4 of 80 μ L can be added（TRIS）Reaction 20min or
The pH value 8.0, ethanolamine concentration of 80 μ L is the phosphate buffer closing magnetic bead of 40mmol/L；With 80 μ L containing 0.15~
The magnetic bead that the phosphate buffer cleaning of the Tween-20 of BSA, 0.15wt.% of 0.22wt.% has been closed 3~4 times, magnetic bead is answered
Be dissolved in containing 0.15~0.22% BSA, 0.015~0.085% Tween-20,0.025wt.% NaN3Phosphate buffer in,
In 2~8 DEG C of preservations.
Embodiment 2：The establishment of kit
The detection kit of sulfamethazine in food is set up, it is contained following component：
The label of the sulfamethazine haptens of horseradish peroxidase-labeled
Enzyme-labelled antigen dilution
The conjugate of sulfamethazine monoclonal antibody and magnetic bead
Magnetic labeling antibody dilution
Sulfamethazine serial standards solution, concentration is respectively：0μg/L、0.025μg/L、0.05μg/L、0.1μg/L、
0.2 μ g/L, 0.4 μ g/L, 0.8 μ g/L, 1.6 μ g/L, standard dilutions are Tween-20 containing 0.035wt.%, 0.042mol/L
The phosphate buffer of pH value 7.4.
It is that liquid, the specifically every liter NaH containing 45~60g are redissolved in 20 times of concentrations that liquid is redissolved in concentration2PO4·2H2O, 200~
220g Na2HPO4·12H2The aqueous solution of O.
Concentrated cleaning solution is 20 times of concentrated cleaning solutions, specifically containing 0.38~0.42wt.% Tween-20s, 2.6~3mol/L
PH value 7.2~7.4 phosphate buffer.
Embodiment 3：The detection of sulfamethazine residual quantity in sample
1st, sample-pretreating method
（1）Milk
Take 25 μ L fresh milks samples and add 950 μ L sample dilutions（To be concentrated with deionized water and redissolve the body that liquid is diluted to 10 times
Product）, whirling motion mixing, taking the solution is used for sample analysis.
（2）Milk powder
0.5g ± 0.05g powdered milk samples are weighed, 5ml sample diluting liquids are added, whirling motion mixes, is taken out 200 μ L and adds to 600
In μ L sample dilutions, whirling motion is mixed, and takes the solution for sample analysis.
2nd, detected and interpretation of result with kit
By enzyme-labelled antigen and enzyme-labelled antigen dilution according to 1：20 volume ratio is diluted, and obtains enzyme-labelled antigen working solution；Will
Magnetic labeling antibody is with magnetic labeling antibody dilution according to 1：20 volume ratio is diluted, and obtains magnetic labeling antibody working solution；30 are taken respectively
~35 μ L enzyme-labelled antigens working solutions, 30~35 μ L samples extract solutions and 30~35 μ L magnetic labeling antibody working solutions, are added sequentially to hold
In device, 18min is reacted at room temperature, then carry out 2~3min of Magneto separate, after abandoning supernatant, with the μ L of cleaning solution 500~800 to multiple
Compound precipitation cleaning 3~4 times；Chemical luminous substrate A liquid and chemical luminous substrate B liquid each 30 are added toward the compound of separator well
μ L, the relative light intensity that detection sends（RLU）, the content of sulfamethazine and RLU, can be with into negative correlativing relation in sample
The residual concentration of sulfamethazine is calculated by RLU standard curves.
The present invention uses 8 sulfamethazine serial standards（0μg/L、0.025μg/L、0.05μg/L、0.1μg/
L、0.2μg/L、0.4μg/L、0.8μg/L、1.6μg/L）Carry out plotting curves.The standard items that to be obtained and sample RLU values
Average value is divided by first RLU value of standard items（RLU0Value）Multiplied by with 100, with relative luminous intensity（%）=RLU/RLU0It is vertical
Coordinate, the logarithm of sulfamethazine concentration does standard curve for abscissa, and the concentration of each sample can be bent from standard
Read on line.
Embodiment 4：The measure of kit quality
1st, the test limit of kit
The definition of kit test limit is：20 negative samples are determined, the average value of measure adds 3 times of standard deviations.The kit
Detection be limited to：The μ g/L of milk 0.91, the μ g/kg of milk powder 0.97.
2nd, the degree of accuracy of kit and precision
The degree of accuracy refers to the matching degree between measured value and true value, and the kit degree of accuracy is often represented with the rate of recovery.Precision also known as
Repeatability, the conventional coefficient of variation is represented.
According to the sample-pretreating method of embodiment 3, with 0.025 μ g/L, 0.05 μ g/L and the sulfanilamide (SN) two of 0.1 μ g/L concentration
Methylpyrimidine is added to milk sample, with 0.05 μ g/kg, 0.1 μ g/kg and the sulfamethazine of 0.2 μ g/kg concentration
Powdered milk sample is added, every kind of sample each concentration mensuration 5 is parallel, is measured with three batches of kits, calculates sample
The rate of recovery and precision.Experimental result shows that the TIANZHU XINGNAO Capsul scope of sulfamethazine is 86.2 in milk sample
~111.7%, the TIANZHU XINGNAO Capsul scope of sulfamethazine is 83.9~108.4% in powdered milk sample.With batch anaplasia in batch
Different coefficient is respectively less than 10%.
3rd, specificity
Using sulfamethazine as standard, if the cross reacting rate of sulfamethazine is 100%, intersect for antibody
The medicine for reacting Journal of Sex Research is and sulfamethazine structure or intimate competition medicine：Sulphadiazine aspergillus flavus
Toxin B1, sulfamethoxazole ochratoxin, 5-methoxysulfadiazine zearalenone.By kit step operation, make
Suppression curve, 50% inhibition concentration of each medicine is calculated according to linear equation（IC50）.Cross reacting rate（%CR）As antibody is to sulphur
The IC of amine dimethyl pyrimidine50With antibody to the IC of sulfamethazine competitor50The ratio between percentage, as a result show：Reagent
Box has specificity higher to sulfamethazine, pair with sulfamethazine structure or intimate strive medicine unexpectedly
The equal no cross reaction of thing.
It is obvious to a person skilled in the art that the invention is not restricted to the details of above-mentioned one exemplary embodiment, Er Qie
In the case of without departing substantially from spirit or essential attributes of the invention, the present invention can be in other specific forms realized.Therefore, no matter
From the point of view of which point, embodiment all should be regarded as exemplary, and be nonrestrictive, the scope of the present invention is by appended power
Profit requires to be limited rather than described above, it is intended that all in the implication and scope of the equivalency of claim by falling
Change is included in the present invention.
Moreover, it will be appreciated that although the present specification is described in terms of embodiments, not each implementation method is only wrapped
Containing an independent technical scheme, this narrating mode of specification is only that for clarity, those skilled in the art should
Specification an as entirety, the technical scheme in each embodiment can also be formed into those skilled in the art through appropriately combined
May be appreciated other embodiment.
Claims (10)
1. in a kind of food sulfamethazine detection kit, it is characterised in that including：Enzyme-labelled antigen, enzyme-labelled antigen
Dilution, magnetic labeling antibody, magnetic labeling antibody dilution, sulfamethazine serial standards solution, chemical luminous substrate A liquid,
Liquid, concentrated cleaning solution are redissolved in chemical luminous substrate B liquid, concentration.
2. in food according to claim 1 sulfamethazine detection kit, it is characterised in that described enzyme
Mark antigen is the label of the sulfamethazine haptens of horseradish peroxidase-labeled, described sulfamethazine
Haptens is to be reacted in methanol solution by sulfamethazine and ethylenediamine and obtained.
3. in food according to claim 1 sulfamethazine detection kit, it is characterised in that described enzyme
Mark antigenic dilution is Na3PO4Concentration is 0.014mol/L, NaCl concentration for the buffering of the pH value 7.2~7.4 of 0.20mol/L is molten
Liquid.
4. in food according to claim 1 sulfamethazine detection kit, it is characterised in that described magnetic
Labeling antibody is that sulfamethazine monoclonal antibody is obtained with the coupling of silicon substrate magnetic bead；Described silicon substrate magnetic bead surfaces group contains
Amount 0.l~0.3eq/g, described magnetic labeling antibody is stored in the Tween-20 containing 0.35~0.4wt.%, 0.035~0.04mol/L
PH value 7.2~7.4 phosphate buffer in.
5. in food according to claim 1 sulfamethazine detection kit, it is characterised in that described magnetic
Labeling antibody dilution is Na2HPO4Concentration is 0.012mol/L, the buffering of the pH value 7.2~7.4 that NaCl concentration is 0.25mol/L
Solution.
6. in food according to claim 1 sulfamethazine detection kit, it is characterised in that described sulphur
Amine dimethyl pyrimidine monoclonal antibody is the conjugate conduct obtained by sulfamethazine haptens and chicken ovalbumin
Balb/c mouse prepare immunogen immune.
7. in food according to claim 1 sulfamethazine detection kit, it is characterised in that described change
Luminous substrate A liquid is luminol content for 0.026~0.028 μ g/L, p-cresol content are 0.04~0.0042 μ g/L, pH
The tris solution of value 8.4~8.6, described chemical luminous substrate B liquid is that every 100ml aqueous solution contains citric acid
2~2.2g of sodium, anhydrous Na2HPO43~3.5g and volumn concentration are 0.65% CO (NH2)2·H2O21.5~1.8ml.
8. in food according to claim 1 sulfamethazine detection kit, it is characterised in that described sulphur
Amine dimethyl pyrimidine serial standards solution concentration is respectively：0μg/L、0.025μg/L、0.05μg/L、0.1μg/L、0.2μg/
L, 0.4 μ g/L, 0.8 μ g/L, 1.6 μ g/L, standard dilutions are the pH value of Tween-20 containing 0.035wt.%, 0.042mol/L
7.4 phosphate buffer.
9. in food according to claim 1 sulfamethazine detection kit, it is characterised in that described is dense
It is that liquid, the specifically every liter NaH containing 45~60g are redissolved in 20 times of concentrations that liquid is redissolved in contracting2PO4·2H2O, 200~220g Na2HPO4·
12H2The aqueous solution of O.
10. in food according to claim 1 sulfamethazine detection kit, it is characterised in that it is described
Concentrated cleaning solution is 20 times of concentrated cleaning solutions, the pH value 7.2 specifically containing 0.38~0.42wt.% Tween-20s, 2.6~3mol/L
~7.4 phosphate buffer.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201611027949.6A CN106771132A (en) | 2016-11-22 | 2016-11-22 | The detection kit of sulfamethazine in a kind of food |
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| CN201611027949.6A CN106771132A (en) | 2016-11-22 | 2016-11-22 | The detection kit of sulfamethazine in a kind of food |
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| CN201611027949.6A Pending CN106771132A (en) | 2016-11-22 | 2016-11-22 | The detection kit of sulfamethazine in a kind of food |
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| CN (1) | CN106771132A (en) |
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| CN106671130A (en) * | 2016-11-28 | 2017-05-17 | 广西大学 | Double-servo-drive multi-rod mechanical arm for assembling work |
| CN107247135A (en) * | 2017-08-11 | 2017-10-13 | 太原瑞盛生物科技有限公司 | A kind of chemiluminescence detection kit of sulfamethazine and preparation method thereof |
| CN108982841A (en) * | 2018-09-25 | 2018-12-11 | 沭阳康源泰博生物科技有限公司 | A kind of sulfamido Sample pretreatment kit based on immunomagnetic beads |
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| CN101936983A (en) * | 2010-08-03 | 2011-01-05 | 中国农业大学 | Method for detecting sulfonamide compound and special quantum dot fluorescent immune kit thereof |
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| CN106671130A (en) * | 2016-11-28 | 2017-05-17 | 广西大学 | Double-servo-drive multi-rod mechanical arm for assembling work |
| CN107247135A (en) * | 2017-08-11 | 2017-10-13 | 太原瑞盛生物科技有限公司 | A kind of chemiluminescence detection kit of sulfamethazine and preparation method thereof |
| CN108982841A (en) * | 2018-09-25 | 2018-12-11 | 沭阳康源泰博生物科技有限公司 | A kind of sulfamido Sample pretreatment kit based on immunomagnetic beads |
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Application publication date: 20170531 |