CN107561269A

CN107561269A - The magnetic microparticle chemiluminescence detection kit and preparation method of a kind of salbutamol

Info

Publication number: CN107561269A
Application number: CN201710723392.8A
Authority: CN
Inventors: 常燕; 胡雪婷; 曹晶; 刘丽青; 杜爱铭; 徐兵
Original assignee: Taiyuan Rui Sheng Biotechnology Co Ltd
Current assignee: Taiyuan Rui Sheng Biotechnology Co Ltd
Priority date: 2017-08-23
Filing date: 2017-08-23
Publication date: 2018-01-09

Abstract

The invention discloses the magnetic microparticle chemiluminescence detection kit and preparation method of a kind of salbutamol.The kit includes：Acridinium ester label, coupling have magnetic particle, salbutamol calibration object, chemiluminescence preexciting liquid A, chemiluminescence exciting liquid B and the cleaning fluid of antigen or antibody.Kit of the present invention is using Magneto separate chemiluminescence as detection means, in combination with acridinium ester label technology.Direct chemoluminescence method high sensitivity that the present invention establishes, high specificity, accurate quick, detection time is short, and testing result has higher accuracy and repeatability, and the kit is applicable to various luminometer devices.

Description

The magnetic microparticle chemiluminescence detection kit and preparation method of a kind of salbutamol

Technical field

The invention belongs to field of detection of food safety, the immune magnetic microparticle chemiluminescence detection of specifically a kind of salbutamol Kit and preparation method.

Background technology

Salbutamol（Salbutamol, SAL）, Chinese nickname is 4- (2- (tert-butylamino) -1- hydroxyethyls) -2- (methylol) phenol, molecular formula C₁₃H₂₁NO₃, its structure is：

Also known as albuterol, albuterol, Controlled Release Salbutamol, salbutamol, Chuan Le Ning, broncovaleas, the one kind clinically commonly used Antiasthmatic, belong to selective β₂Receptor stimulating agent, have to animal and promote skeletal muscle（Lean meat）Growth, lipopexia is reduced, improved Feed conversion rate etc. is acted on, and the production of livestock products is often illegally used for as feed addictive.Salbutamol is survivable, if Animal is not by the medication retention of certain time before butchering, and residual can be gathered in animal edible tissues, due to this Class compound has Orally active, if illegal Use out of range people know from experience will appear from different degrees of toxic reaction, its symptom with Animal poisoning symptom is similar, shows as muscular tremor, quadriplegia, tachycardia, arrhythmia cordis, stomachache, myalgia, nausea The symptoms such as dizziness, expiratory dyspnea, severe one can trigger hypertension, heart disease even dead, have a strong impact on health.Therefore include Many countries all successively legislations including China prohibit the use of the salbutamol for promoting growth of animal as feed addictive.

Early in March, 1997, application of the beta-2-agonists in animal productiong is forbidded strictly in Ministry of Agriculture's dispatch.2001, with The ground such as Guangdong, Guangxi and recur the edible animal food poisoning containing beta-2-agonists of people, the Ministry of Agriculture starts " no public affairs Evil food action plan ", by emphasis of " clenbuterol hydrochloride " monitoring as animal product monitoring.To strengthen residue of veterinary drug monitoring work, Ensure that animal food is safe and healthy, China is in No. 235 bulletin of the Ministry of Agriculture《Animal food herbal medicine MRL》 Middle clear stipulaties salbutamol and its salt, ester are the medicine prohibitted the use of, must not be detected in animal food.Therefore, eat Salbutamolum residue detects in product, to ensureing that food security plays vital effect.

Up to the present, the method for detecting salbutamolum residue in animal derived food mainly has：High-efficient liquid phase color Spectrometry（HPLC）, gas chromatography-mass spectrography（GC-MS）, liquid chromatograph mass spectrography（LC-MS）, colloidal gold immunity chromatography （Colloidal gold immunochromatography）, ELISA（ELISA）, enzyme-catalyzed chemical luminescence etc..But High performance liquid chromatography, gas chromatography-mass spectrography, Liquid Chromatography-Mass Spectrometry Instrumental equipment are expensive, sample pre-treatments Complexity, waste time and energy and be not easy to popularize, testing cost is high.CN 103018452 A（In April, 2013）It is husky to disclose a kind of detection The colloidal gold test paper card and detection method of butylamine alcohol medicine, salbutamol-carrier protein is coated with the reaction film of the kit The detection line and be coated with the nature controlling line that sheep anti mouse antiantibody is formed that conjugate is formed, conjugate release pad is coated with salbutamol Monoclonal antibody-colloid gold label thing, then use the conjugate release pad, reaction film and the sample absorbent that prepare, adsorptive pads Test card is assembled into backing, is finally detected with colloidal gold test paper card, the major defect of this method is cumbersome, flow It is more, it is more easy to error occur, sensitivity is low.Although ELISA detection is cheap, quick, sensitivity is also inadequate, only Suitable for the detection and identification of micro substance, the A of CN 103018447（In April, 2013）Disclose a kind of detection salbutamol Enzyme linked immunological kit and its method, the kit is using ELISA Plate coating salbutamol drug antigenic, horseradish peroxidase Salbutamol mouse monoclonal antibody is marked, is added after the two is mixed in sample, substrate solution is then added and is developed the color, measure is inhaled For shading value so as to calculate the concentration of salbutamol in sample, the major defect of this method is that sensitivity is low.CN 105315241 A（2 months 2016）Disclose a kind of salbutamol haptens and antigen and its chemiluminescence immunoassay kit special, the kit Using polystyrene Chemiluminescent plate coating salbutamol and artificial antigen, horseradish peroxidase made of carrier protein coupling Rabbit source polyclonal antibody is marked, sample to be tested solution is added in microwell plate, then adds primary antibody and ELIAS secondary antibody, sample to be tested Coated coating antigen competition primary antibody, then further with ELIAS secondary antibody on the salbutamol and Chemiluminescent plate remained in solution With reference to then adding Chemoluminescent substrate, the content of salbutamol in sample calculated by luminous intensity values, using horseradish Peroxidase major defect is：Luminol, also can be by H in the case of the presence of no horseradish peroxidase₂O₂Aoxidize itself hair Light, background is of a relatively high, influences signal to noise ratio, kinetics is complicated, and influence factor is more, is as a result not sufficiently stable, and to obtain sensitive Degree is high and the substrate of plateau length is not easy.Therefore, a kind of detection effective, quick, simple, sensitive, anti-interference is high is established The method tool of salbutamol is of great significance.

The present invention uses method as direct chemoluminescence method, using acridinium ester as chemiluminescent labels with obvious excellent Gesture, it is mainly manifested in：Reaction does not need catalyst, as long as alkaline environment can be carried out, is swift in response, can complete catching reaction Caused photon, background luminescence is low, and signal to noise ratio is high, and disturbing factor is few, and reagent stability is good, and system is simple, and exciting liquid cost is low, Easily and protein bind, and photon yield is not reduced acridinium ester after being coupled.The Magnetism particulate immuno chemistry luminescence method that the present invention establishes is sensitive Spend height, high specificity, accurate quick, detection time is short, testing result have higher accuracy with it is repeated.

The content of the invention

It is an object of the invention to provide the sand that a kind of sensitivity is higher, the reaction time is short, simple to operate, anti-interference is high The magnetic microparticle chemiluminescence detection kit and preparation method of butylamine alcohol.

To achieve the above object, the present invention provides following technical scheme：

The magnetic microparticle chemiluminescence detection kit and preparation method of a kind of salbutamol, magnetic particle chemistry provided by the present invention Luminescent method detects the kit of salbutamol, and salbutamol monoclonal antibody can be taken to be coupled magnetic particle, acridinium ester label Salbutamol antigen, salbutamol antigen can also be taken to be coupled magnetic particle, acridinium ester label salbutamol monoclonal antibody.Examination Agent box also include salbutamol calibration object, above-mentioned acridinium ester act on chemiluminescence preexciting liquid A, chemiluminescence exciting liquid B with And cleaning fluid.

Described magnetic particle directly can be coupled with antibody or antigen, can be also coupled magnetic particle and Streptavidin, simultaneously Using biotin labelled antibodies or antigen.

The surface modification group of the magnetic particle is carboxyl, amino, p-toluenesulfonyl or Streptavidin-biotin.

Described chemiluminescent labels are acridinium ester, such as NSP-DMAE-NHS, NSP-SA-NHS.

Described acridinium ester label is salbutamol antigen, or salbutamol monoclonal antibody.

Described calibration object is to be with the Tris-HCl buffer solutions containing 0.5-5.0% BSA and 0.1-0.5% PC300 Matrix, add the configuration of salbutamol sterling and form, calibration object form is liquid.

Described salbutamol calibration object solution concentration is respectively：0 μg/L、0.01 μg/L、0.1 μg/L、1.0 μg/ L、10.0 μg/L、20.0 μg/L。

Described chemiluminescence preexciting liquid A is H₂O₂And HNO₃Mixed liquor, wherein H₂O₂Mass fraction be 0.05- 5%, HNO₃Concentration be 0.05-2.5 mol/L.

Described chemiluminescence exciting liquid B is Triton X-100 and NaOH mixed liquor, wherein Triton X-100's Concentration is that 0.05-2.0 mol/L, NaOH concentration are 0.05-1.0 mol/L.

Described cleaning fluid is：PH 7.0-9.0, the Tris-HCl solution that concentration is 5.0-50.0 mmol/L, wherein containing Concentration is 0.05-0.50 mol/L NaCl and 0.01-0.25% Tween-20.

The principle of the present invention is to be combined the high degree of specificity of antibody-antigene reaction with the high sensitivity that acridinium ester lights Get up, using photon caused by acridinium ester catching reaction to detect production concentration.

The advantage of the invention is that combining magnetic microparticle chemiluminescence technology using competition law, the salbutamol in food is determined Content.Acridinium ester has a clear superiority as the direct chemiluminescence of label, is mainly manifested in：Reaction does not need catalyst, As long as alkaline environment can be carried out, be swift in response, can photon completely caused by catching reaction, background luminescence is low, and signal to noise ratio is high, Disturbing factor is few, and reagent stability is good, and system is simple, and exciting liquid cost is low, acridinium ester easily and protein bind, and light after being coupled Sub- yield is not reduced.

Embodiment

The present invention provides a kind of the magnetic microparticle chemiluminescence detection kit and preparation method of salbutamol, to make the present invention Purpose, technical scheme and effect definitely, it is clear, the present invention is described in more detail below.

The present invention provides a kind of the magnetic microparticle chemiluminescence detection kit and preparation method of salbutamol, wherein, this hair The kit of bright provided magnetic microparticle chemiluminescence method detection salbutamol, can take salbutamol monoclonal antibody even Join magnetic particle, acridinium ester label salbutamol antigen can also take salbutamol antigen to be coupled magnetic particle, and acridinium ester label is husky Butylamine alcohol monoclonal antibody.The chemiluminescence preexciting liquid that kit also includes salbutamol calibration object, above-mentioned acridinium ester acts on A, chemiluminescence exciting liquid B and cleaning fluid.

Specifically, the kernel of magnetic bead of the present invention is ferroso-ferric oxide, described magnetic particle can directly with antibody or anti- Original coupling, magnetic particle and Streptavidin can be also coupled, while use biotin labelled antibodies or antigen.It is solid before magnetic bead use The endless bulk deposition of phase can influence accuracy, therefore should select good dispersion when selecting, and it is few to place magnetic bead number of uniting for a long time, sinks Slow-footed magnetic bead drops.

Specifically, for the present invention when preparing magnetic particle suspension, the coupled antigen buffer solution is pH 5.0, concentration is 0.1 mol/L MES buffer solutions；The MES buffer solutions that coupled antibody buffer solution is pH 6.0, concentration is 0.1 mol/L.

Specifically, for the present invention when preparing magnetic particle suspension, the Block buffer is the buffer solution containing 1%BSA.

Specifically, calibration object of the present invention is with the Tris-HCl containing 1-3% BSA and 0.1-0.3% PC300 Buffer solution is matrix, adds the configuration of salbutamol sterling and forms, calibration object form is liquid.

Specifically, chemiluminescence preexciting liquid A of the present invention is H₂O₂And HNO₃Mixed liquor, wherein H₂O₂Quality Fraction is 1.5%, HNO₃Concentration be 0.1 mol/L.

Specifically, chemiluminescence exciting liquid B of the present invention is Triton X-100 and NaOH mixed liquor, wherein Triton X-100 concentration is 0.1 mol/L, and NaOH concentration is 0.35 mol/L.

Specifically, cleaning fluid of the present invention is：PH 7.2, the Tris-HCl solution that concentration is 25 mmol/L, wherein containing Concentration is 0.15 mol/L NaCl and 0.05% Tween-20.

Below by embodiment, the present invention is described in detail.

Embodiment 1：A kind of establishment of the magnetic microparticle chemiluminescence kit 1 of described detection salbutamol and preparation side Method, comprise the following steps：

1. the establishment of kit 1：

A kind of magnetic microparticle chemiluminescence kit for detecting salbutamol is set up, it is contained following component：

The salbutamol monoclonal antibody of carboxyl magnetic particle coupling；

The salbutamol antigen of acridinium ester label；

Salbutamol serial standards solution, concentration are respectively：0 μg/L、0.01 μg/L、0.1 μg/L、1.0 μg/L、10.0 μ g/L, 20.0 μ g/L, its buffer solution are the Tris-HCl solution containing 1-3% BSA and 0.1-0.3% PC300；

Chemiluminescence preexciting liquid A and chemiluminescence exciting liquid B；

The Tris-HCl buffer solutions of cleaning fluid, the specially mmol/L of concentration 25（pH 7.2）, wherein containing the mol/L of concentration 0.15 NaCl and 0.05% Tween-20.

2. coupling has the preparation of the magnetic particle suspension of salbutamol monoclonal antibody

（1）1 mg carboxyls magnetic particle is taken in 0.5 mL centrifuge tubes, adds a certain amount of 0.1 mol/L MES buffer solutions, is vortexed mixed It is even, it is placed on magnetic frame, standing 5 min makes magnetic particle be separated with liquid, supernatant discarding, washs 3 times, adds a certain amount of MES （PH is 6.0）Buffer solution, it is vortexed.

（2）Add 15 μ L（15 μg）Salbutamol monoclonal antibody, be vortexed, revolving reaction pipe, incubation at room temperature 17 min。

（3）Add the coupling reagent EDC that 10 μ L concentration are 10 mg/mL to be vortexed, revolving reaction pipe, be incubated at room temperature 2 h.

（4）Supernatant is removed, adds a certain amount of cleaning buffer solution（TBS+0.05%Tween-20）, wash 3 times.

（5）Closed with the buffer solution containing 1%BSA, repeatedly closing 4 times, every time 10 min.By the magnetic particle suspension It is placed in 2-8 DEG C of preservation.

3. prepared by the salbutamol antigen liquid-phase reagent of acridinium ester label

（1）Purify salbutamol：A certain amount of salbutamol antigen is placed in bag filter, and bag filter is placed in not less than 1 Dialysed in L mark buffer solution, during which buffer solution is at least changed 3 times, last time dialysed overnight, and mark buffer solution is pH 10.1st, concentration is 0.1 mol/L Na₂CO₃-NaHCO₃Buffer solution.

（2）1.7 mg acridinium ester NSP-DMAE-NHS is weighed, is dissolved in 447 μ L anhydrous dimethyl formamides DMF, matches somebody with somebody Into 6.5 mmol/L NSP-DMAE-NHS DMF solutions.

（3）It will be placed in through the albuterol solution after dialysis in 500 μ L centrifuge tubes（Lucifuge is reacted）, add 200 μ L Buffer solution is marked, then adds a certain amount of 6.5 mmol/L NSP-DMAE-NHS DMF solutions, acridinium ester and salbutamol Molar ratio be 9.7:1,1 h is reacted at room temperature, adds the μ L of 10 g/L lysines 100, is continued to react 15 min, is made mark Reaction terminating.

（4）Label NSP-DMAE-NHS-Ag and free NSP-DMAE-NHS passes through Sephadex G-50 posts（1× 25cm）Separation, the PBS for being 0.1mol/L with purification buffer pH 6.3, concentration are balanced and are eluted chromatographic column.

（5）Protein peak is detected with chromatograph in separation process, the chemiluminescence intensity and 280nm for measuring efflux respectively are inhaled Shading value.

（6）The eluent that shading value is high and absorbance is big is collected, adds 1% BSA（Volume）After dispense.

4. salbutamol calibration object is prepared

Salbutamol sterling is configured to indicate with the Tris-HCl buffer solutions containing 1-3% BSA and 0.1-0.3% PC300 Concentration is 0 μ g/L, 0.01 μ g/L, 0.1 μ g/L, 1.0 μ g/L, 10.0 μ g/L, the calibration object of 20.0 μ g/L totally 6 concentration Grad.

5. prepared by chemiluminescence exciting liquid A, B

（1）Chemiluminescence preexciting liquid A is by H₂O₂And HNO₃Composition.Wherein, wherein H₂O₂Mass fraction be 1.5%, HNO₃It is dense Spend for 0.1 mol/L, be distributed into 20 mL/ branch with brown bottle, 2-8 DEG C saves backup.

（2）Chemiluminescence exciting liquid B is made up of Triton X-100 and NaOH mixed liquor.Wherein, Triton X-100 Concentration be 0.1 mol/L, NaOH concentration is 0.35 mol/L, is distributed into 20 mL/ branch with brown bottle, 2-8 DEG C of preservation is standby With.

Embodiment 2：A kind of establishment of the magnetic microparticle chemiluminescence kit 2 of described detection salbutamol and preparation side Method, comprise the following steps：

1. the establishment of kit 2：

The salbutamol antigen of carboxyl magnetic particle coupling；

The salbutamol monoclonal antibody of acridinium ester label；

2. coupling has the preparation of the magnetic particle suspension of salbutamol antigen

（1）1 mg carboxyls magnetic particle is taken in 0.5 mL centrifuge tubes, adds a certain amount of 0.1 mol/L MES buffer solutions, is vortexed mixed It is even, it is placed on magnetic frame, standing 5 min makes magnetic particle be separated with liquid, supernatant discarding, washs 3 times, adds a certain amount of MES （PH is 5.0）Buffer solution, it is vortexed.

（2）Add 15 μ L（15 μg）Salbutamol antigen, be vortexed, revolving reaction pipe, be incubated at room temperature 17 min.

（4）Supernatant is removed, adds 200 μ L cleaning buffer solution（TBS+0.05% Tween-20）, wash 3 times.

3. prepared by the salbutamol monoclonal antibody liquid-phase reagent of acridinium ester label

（1）Purify salbutamol monoclonal antibody：A certain amount of salbutamol monoclonal antibody is placed in bag filter, and will be saturating Analysis bag, which is placed in the mark buffer solution not less than 1 L, is dialysed, and during which buffer solution is at least changed 3 times, last time dialysed overnight, mark The Na that note buffer solution is pH 10.1, concentration is 0.1 mol/L₂CO₃-NaHCO₃Buffer solution.

（3）It will be placed in through the salbutamol monoclonal antibody solution after dialysis in 500 μ L centrifuge tubes（Lucifuge is reacted）, 200 μ L mark buffer solutions are added, then add a certain amount of 6.5 mmol/L NSP-DMAE-NHS DMF solutions, acridinium ester Molar ratio with salbutamol monoclonal antibody is 7.4:1,1h is reacted at room temperature, adds the μ L of 10 g/L lysines 100, after 15 min of continuous reaction, make mark reaction terminating.

（4）Label NSP-DMAE-NHS-Ab and free NSP-DMAE-NHS passes through Sephadex G-50 posts（1× 25cm）Separation, balanced with purification buffer pH 6.3, the PBS that concentration is 0.1 mol/L and elute chromatographic column.

4th, salbutamol calibration object is prepared

5. prepared by chemiluminescence exciting liquid A, B

（1）Chemiluminescence preexciting liquid A is by H₂O₂And HNO₃Composition.Wherein, H₂O₂Mass fraction be 1.5%, HNO₃Concentration be 0.1 mol/L, 20 mL/ branch are distributed into brown bottle, 2-8 DEG C saves backup.

Embodiment 3：The pre-treatment of sample

A. the processing of muscle, internal organ

The processing of muscle：The tissue samples after 2.0 g homogeneous are weighed into 50 mL centrifuge tubes；Add 4 mL, 2% NaCl and 0.2 mol/L HCl- methyl alcohol mixed liquors, vibrate 30 s；0.5 mL supernatants are taken (to keep away after 3000 r/min, room temperature centrifugation 5min Open upper strata suspension) 35 mL, 0.5 mol/L NaOH solutions are added, add 0.5 mL, the phosphoric acid that concentration is 0.02 mol/L Salt buffer, mix.

The processing of internal organ：The tissue samples after 2.0 g homogeneous are weighed into 50 mL centrifuge tubes；Add 4 mL, 2%NaCl with And 0.2 mol/L HCl- methyl alcohol mixed liquor, vibrate 30 s；3000 r/min, room temperature take 0.5 mL supernatants after centrifuging 5 min (avoiding upper strata suspension) adds 20 mL, 0.5 mol/L NaOH solutions, adds 0.5 mL, concentration is 0.02 mol/L's Phosphate buffer, mix.

B. the processing of milk test sample solution

150 μ L fresh milks are taken in 500 μ L centrifuge tubes, 4 DEG C of 10 min of centrifugation（3000 r/min）, discard upper-layer fat. Pipette the μ L of milk sample 25 after centrifugation to be placed in clean teat glass, add 950 μ L, concentration is 0.02 mol/L phosphoric acid Salt buffer is diluted.

Embodiment 4：The step of being detected using the magnetic microparticle chemiluminescence detection kit of described salbutamol be：

（1）By μ L of sample to be tested 100, μ L of coupling magnetic particle suspension 150, the μ L of acridinium ester label 150, sequentially add anti- Ying Guanzhong, vibration mix, 37 DEG C of 15 min of incubation.

（2）Separation cleaning 5 times.

（3）Reaction vessel after washing, which is fully vibrated, makes magnetic particle scatter.

（4）100 μ L chemiluminescence exciting liquid B are added after adding 100 μ L chemiluminescence preexciting liquid A, 1 s, measure it Relative luminous intensity, the content of salbutamol luminous intensity proportion relation corresponding thereto in sample.

Embodiment 5：The performance indications of kit

（1）The sensitivity of kit

20 retests are carried out to zero standard solution, the average value for taking zero standard solution to determine adds 3 times of standard deviation, i.e., For the sensitivity of this kit.Sensitivity of this kit to salbutamol is 0.02 μ g/L.

（2）The specificity of kit

The competition medicine similar to salbutamol structure or function：Ractopamine, Clenbuterol, Terbutaline, bromine Boot sieve, Phenolethanolamine A.By kit step operation, salbutamol, Ractopamine, Clenbuterol, Terbutaline, bromine cloth are separately added into Special sieve, phenolethanolamine A, suppression curve is made, 50% inhibition concentration of each medicine is calculated according to linear equation.Cross reacting rate is IC for antibody to salbutamol₅₀IC with antibody to salbutamol competitor₅₀The ratio between percentage.As a result show：This reagent Box has higher specificity to salbutamol, pair with salbutamol structure or it is intimate competition medicine without intersects instead Should.

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 realized in other specific forms.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 rather than described above limits, 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 embodiment is only wrapped Containing an independent technical scheme, this narrating mode of specification is only that those skilled in the art should for clarity Using specification as an entirety, the technical solutions in the various embodiments may also be suitably combined, forms those skilled in the art It is appreciated that other embodiment.

Claims

1. the magnetic microparticle chemiluminescence detection kit and its composition of a kind of salbutamol, it is characterised in that：There is sand including coupling The magnetic particle suspension of butylamine alcohol monoclonal antibody, calibration object, the salbutamol antigen of acridinium ester label, chemiluminescence preexciting Liquid A, chemiluminescence exciting liquid B and cleaning fluid.

2. the magnetic microparticle chemiluminescence detection kit and its composition of a kind of salbutamol according to claim 1, it is special Sign is：Described magnetic particle directly can be coupled with salbutamol monoclonal antibody, or magnetic particle and Streptavidin are coupled, Use biotin labeling salbutamol monoclonal antibody simultaneously.

3. the magnetic microparticle chemiluminescence detection kit and its composition of a kind of salbutamol according to claim 1, it is special Sign is：Described chemiluminescent labels are acridinium ester, such as NSP-DMAE-NHS, NSP-SA-NHS.

4. the magnetic microparticle chemiluminescence detection kit and its composition of a kind of salbutamol according to claim 1, it is special Sign is：Described acridinium ester label is salbutamol antigen.

5. the magnetic microparticle chemiluminescence detection kit and its composition of a kind of salbutamol according to claim 1, it is special Sign is：Described calibration object be using the Tris-HCl buffer solutions containing 0.5-5.0% BSA and 0.1-0.5% PC300 as Matrix, add the calibration object solution for the series concentration gradient that the configuration of salbutamol sterling forms.

6. the magnetic microparticle chemiluminescence detection kit and its composition of a kind of salbutamol according to claim 1, it is special Sign is：Described chemiluminescence preexciting liquid A is by H₂O₂And HNO₃Mixed liquor composition, chemiluminescence exciting liquid B is by Triton X-100 and NaOH mixed liquor composition.

7. the magnetic microparticle chemiluminescence detection kit and its composition of a kind of salbutamol, it is characterised in that：There is sand including coupling The magnetic particle suspension of butylamine alcohol antigen, calibration object, the salbutamol monoclonal antibody of acridinium ester label, chemiluminescence preexciting Liquid A, chemiluminescence exciting liquid B and cleaning fluid.

8. the magnetic microparticle chemiluminescence detection kit and its composition of a kind of salbutamol according to claim 7, it is special Sign is：Described magnetic particle directly can be coupled with salbutamol antigen, or magnetic particle and Streptavidin are coupled, and be adopted simultaneously With biotin labeling salbutamol antigen.

9. the magnetic microparticle chemiluminescence detection kit and its composition of a kind of salbutamol according to claim 7, it is special Sign is：Described chemiluminescent labels are acridinium ester, such as NSP-DMAE-NHS, NSP-SA-NHS.

10. the magnetic microparticle chemiluminescence detection kit and its composition of a kind of salbutamol according to claim 7, it is special Sign is：Described acridinium ester label is salbutamol monoclonal antibody.