CN104931689A - Molecularly imprinted test paper strip and preparation method as well as application thereof - Google Patents
Molecularly imprinted test paper strip and preparation method as well as application thereof Download PDFInfo
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Abstract
The invention discloses a clenbuterol hydrochloride molecularly imprinted test paper strip and a preparation method thereof. A nitrocellulose membrane is immerged in a clenbuterol hydrochloride molecularly imprinted polymer solution for soaking; afterwards, the soaked nitrocellulose membrane is oven-dried; the nitrocellulose membrane is washed by using ethyl alcohol to elute template molecules clenbuterol hydrochloride in the imprinted polymer; the washed nitrocellulose membrane is aired; afterwards, the aired nitrocellulose membrane is stuck in a substrate piece with a same hollowed-out area in a pressing manner. By using the test paper strip, the defects that a test paper strip of a same type easily generates a false-negative result and a false-positive result, does not display a quality control line, is too high in detection omission rate, and the like are overcome. A kit prepared on the basis of the test paper strip can be used for carrying out qualitative, semiquantificational and quantificational analysis on the clenbuterol hydrochloride.
Description
Technical field
The present invention relates to molecular imprinting field, be specifically related to a kind of clenobuterol hydrochloride molecular engram test strips and preparation method thereof and application.
Background technology
Clenbuterol hydrochloride is the general designation of a class medicine with adrenocepter agonism, also known as β 2-excitant, there is the effect promoting muscle growth, improve lean meat percentage, Zeng Zuowei growth additive is by extensive concern, but broxaterol has certain residence time in animal body, some Susceptible population can be made to produce obvious toxicity symptom after entering human body by food chain, harm humans is healthy.Clenobuterol hydrochloride (Clenbuterol, CLB) is the one in clenbuterol hydrochloride, and it obviously can promote growth of animal, and increases lean meat percentage, is thus used in animal productiong by some lawless persons and is used for increasing lean meat percentage.Research shows, the organ such as kidney, liver can be poisoned after residual clenobuterol hydrochloride is in animal body used by people, there will be headache discomfort, n and V, muscle tremors, increased heart rate, the symptom such as to have a dizzy spell, even death can be caused, therefore, China forbids to use clenobuterol hydrochloride in animal productiong.The method of existing detection clenobuterol hydrochloride all needs complicated sample handling processes and analytical procedure, and need large-scale instrument and operate through the professional of specialized training, the time is long, cost is high, far can not meet the requirement of on-line checkingi.
Test paper method, owing to having the features such as easy to carry, simple to operate, finding speed is fast, has more application in fields such as food, water quality, health cares.If animal immunology key lab of academy of agricultural sciences of calendar year 2001 Henan Province and Henan Bao'ao Biology Engineering Co., Ltd are in successfully have developed " clenbuterol hydrochloride " special quick detection test paper technology and putting on market, use this test paper whether can detect to quicklook in 60 seconds in meat containing " clenbuterol hydrochloride " that exceed safety standard, this detection paper technology designs based on monoclonal antibody, during detection colloid gold label the tested survey line of " clenbuterol hydrochloride " monoclonal antibody on immobilized " clenbuterol hydrochloride " artificial conjugated antigen catch, form a henna detection line, excessive colloid gold label monoclonal antibody continues to spring up is caught formation henna control line by sheep anti-mouse antibody immobilized on control line, if containing " clenbuterol hydrochloride " in sample, it competes limited colloid gold label monoclonal antibody by with the artificial antigen be immobilized on detection line.Application number is the Chinese patent of 200810219632.1, disclose a kind of clenobuterol hydrochloride Test paper and detection method thereof, this paper bag is drawn together and is supported fixing sticky lining, the fixing sticky lining of support there is sample adsorption layer successively, pad, microporous siphon responding layer and absorbent material layer, the antibody A of antibody of clenbuteral hydrochloride pad having trace particle mark and trace particle mark, when containing the clenobuterol hydrochloride of more than 5 nanograms in sample, there are two lines in detection zone, when in sample, clenobuterol hydrochloride content is less than 5 nanogram, there is a line in detection zone, this test paper has the detecting pattern and interpretation mode that are equal to sandwich method and cost is low, this detection method is simple and efficient, simple to operate.But in use there is following problem in said method: exceeding the interpretation time easily occurs false negative, false positive results, nature controlling line does not manifest sometimes, detection line color is smudgy, loss is higher, therefore, find a kind ofly quick and easyly, high to select, the test paper method of high-sensitive detection clenobuterol hydrochloride is significant.
Molecular engram is the molecular specificity recognition technology of rising in recent years, there is binding site template molecule at functional group, molecular dimension, space structure to memory function, molecular recognition can be carried out according to predetermined selectivity and level identification performance, the specific recognition site energy formed in trace and analyte selectively acting, and this effect is reversible, the mechanical strength of this synthesis and chemical stability good, can use in extreme environment.The now existing molecular engram test strips that molecular imprinting is prepared in conjunction with test paper method, not only possesses molecular engram specific recognition ability, and there is ELISA test strip speed feature fast, easy to carry, but not yet there is the relevant report preparing clenobuterol hydrochloride molecular engram test strips and detection method thereof at present.
Summary of the invention
The present invention is directed to the detection method complex pretreatment of existing classical clenobuterol hydrochloride, the time is long, cost is high, easily there is the present situations such as false negative false positive results, loss are high in existing test paper method, provide a kind of clenobuterol hydrochloride molecular engram test strips and preparation method thereof and application, this test strips preparation method is simple, cost is low, high specificity, good stability.Clenobuterol hydrochloride molecular engram test strips of the present invention can be used for the online trace detection of clenobuterol hydrochloride.
Technical scheme of the present invention is:
A kind of molecular engram test strips, comprises substrate and reaction zone, and described reaction zone is nitrocellulose filter clenobuterol hydrochloride to Selective recognition ability covering substrate void region;
Described preparation method clenobuterol hydrochloride to the nitrocellulose filter of Selective recognition ability is: nitrocellulose filter is put into clenobuterol hydrochloride imprinted polymer solution and soak 30 minutes, take out rearmounted 40 DEG C of drying in oven, with 15-40% ethanol washing nitrocellulose filter, wash nitrocellulose filter with water again, dry, to obtain final product;
Described clenobuterol hydrochloride imprinted polymer solution take clenobuterol hydrochloride as template molecule, take α-methacrylic acid as function monomer, interact with hydrogen bond formation, add crosslinking chemical N, N-methylene-bisacrylamide, initiator ammonium persulfate occur free-radical polymerized, obtain clenobuterol hydrochloride imprinted polymer solution; The mol ratio of described template molecule, function monomer, crosslinking chemical is 117-274:1-300:30-90.
The above molecular engram test strips, described reaction zone can be positioned at any position of substrate, for convenience of operation, is preferably placed at one end of substrate.
The above molecular engram test strips, the material of described substrate can be any material not affecting test, is preferably plastics or photo paper.
The preparation method of the molecular engram test strips described in more than one, comprises the following steps:
1. the preparation of clenobuterol hydrochloride imprinted polymer solution:
A. compound concentration is 3 × 10
-3-3 × 10
-5the clenobuterol hydrochloride aqueous solution of mol/L;
B. 3-7 μ L α-methacrylic acid being joined 10-30mL concentration is 3 × 10
-3-3 × 10
-5in mol/L clenobuterol hydrochloride aqueous solution, ultrasonic process 5-15 minute, then add 0.0014-0.0042g N, N-methylene-bisacrylamide, continue ultrasonic process 10-30 minute, hold over night, obtains mixed liquor;
C. be the ammonium persulfate solution of 0.5mol/L by 80-120 μ L concentration, slowly join in the mixed liquor that step b obtains, ultrasonic process 10-20 minute, namely obtains clenobuterol hydrochloride molecularly imprinted polymer.
2. pair clenobuterol hydrochloride has the preparation of the nitrocellulose filter of Selective recognition ability: nitrocellulose filter is put into clenobuterol hydrochloride imprinted polymer solution and soak 30 minutes, take out rearmounted 40 DEG C of drying in oven, with 15-40% ethanol washing nitrocellulose filter, wash nitrocellulose filter with water again, dry, cut out;
3. cut out substrate, a void region is cut out out in substrate, apply glue at the substratel of void region or be stained with double faced adhesive tape, be bonded in substrate by what cut out in above-mentioned steps 2 to the nitrocellulose filter that clenobuterol hydrochloride has a Selective recognition ability, cover the substrate that pressure one has identical void region again, to obtain final product.
The preparation method of the above molecular engram test strips, in described step 3, the size of substrate is preferably 8cm × 1cm, and the size of described void region is preferably 4cm × 0.8cm.
The application of above-described molecular engram test strips, for clenobuterol hydrochloride in testing sample qualitative, sxemiquantitative or quantitatively detect.Described testing sample can be and anyly may contain clenobuterol hydrochloride material, as feed, urine etc., carries out pre-service, be prepared into testing sample solution by methods such as acid, alkali or enzymolysis.
Above-described molecular engram test strips, for convenient to use, can be made into kit to support the use, this kit comprises molecular engram test strips, active glucose oxidase enzyme labeling clenobuterol hydrochloride solution, 0.1g/mL glucose solution, 3 × 10 for 200-400ug/mL
-4mol/L affixes one's name to red Y solution and standard color comparison card.
The above kit, also comprises purple light pen.
The present invention utilizes chemical method to be polymerized the molecularly imprinted polymer preparing clenobuterol hydrochloride, be prepared into molecular engram test strips, with 25% ethanol elution template molecule clenobuterol hydrochloride, leave trace hole clenobuterol hydrochloride to specific recognition capability.The hole on blotting membrane surface can identify clenobuterol hydrochloride simultaneously and have the clenobuterol hydrochloride of mutually isostructural glucose oxidase enzyme labeling.After the test strips of eluted template molecule being hatched in the clenobuterol hydrochloride of the glucose oxidase enzyme labeling of high concentration, trace hole is occupied by the clenobuterol hydrochloride of glucose oxidase enzyme labeling.Competitive reaction be subsequently glucose oxidase enzyme labeling on clenobuterol hydrochloride in the sample to which and blotting membrane clenobuterol hydrochloride between carry out, clenobuterol hydrochloride the clenobuterol hydrochloride of competitive substitution glucose oxidase enzyme labeling can occupy hole again.Along with the increase of clenobuterol hydrochloride molecular conecentration in sample, more by the clenobuterol hydrochloride of the glucose oxidase enzyme labeling of competitive substitution on blotting membrane, the glucose oxidase namely remained on blotting membrane is fewer.And the glucose oxidase on blotting membrane surface can produce hydrogen peroxide with the glucose generation enzyme digestion reaction in end liquid, the red Y of administration in liquid at the bottom of the further oxygenolysis of hydrogen peroxide makes it fade, therefore, by the fluorescence intensity of test strip, fast qualitative, sxemiquantitative or quantitative test can be carried out to the clenobuterol hydrochloride in sample.
The invention has the beneficial effects as follows:
1. clenobuterol hydrochloride molecular engram test strips of the present invention, is applicable to the fast qualitative of clenobuterol hydrochloride in the sample such as food or excreta, sxemiquantitative or quantitatively detects.
2. test strips preparation method of the present invention is simple, cost is low, high specificity, good stability.
3. clenobuterol hydrochloride molecular engram test strips of the present invention is for detecting clenobuterol hydrochloride, have simple, fast, cost is low, the advantage such as accurately and reliably, avoids occurring false negative false positive results, the situation such as undetected.
4. by test strips generate a reagent box of the present invention, jointly sell, testing process can be made more convenient, realize online trace detection.
Accompanying drawing explanation
Fig. 1 is preparation method's schematic diagram of embodiment of the present invention 1-3 clenobuterol hydrochloride molecular engram test strips.
In figure, each numbering title is respectively: 1. substrate, 2. void region, 3. cementing or sticky double faced adhesive tape region, and 4. pair clenobuterol hydrochloride has the nitrocellulose filter of Selective recognition ability, 5. has the substrate of identical void region, 6. reaction zone
Fig. 2 is the fluorogram of each concentration clenobuterol hydrochloride standard solution of the present invention
The solution that in figure, each numbering test strips is used is respectively: 1. concentration is 3 × 10
-4the clenobuterol hydrochloride standard solution of mol/L, 2. concentration is 3 × 10
-5the clenobuterol hydrochloride standard solution of mol/L, 3. concentration is 3 × 10
-6the clenobuterol hydrochloride standard solution of mol/L, 4. concentration is 3 × 10
-7the clenobuterol hydrochloride standard solution of mol/L, 5. concentration is 3 × 10
-8the clenobuterol hydrochloride standard solution of mol/L, 6. blank solution
Fig. 3 is the fluorogram of the embodiment of the present invention 4 test strips.
Embodiment
Below in conjunction with the drawings and the specific embodiments, the invention will be further described, but do not limit usable range of the present invention and range of application.
One, the preparation method of clenobuterol hydrochloride molecular engram test strips
Embodiment 1
A preparation method for molecular engram test strips, comprises the following steps:
1. the preparation of clenobuterol hydrochloride imprinted polymer solution:
A. compound concentration is 3 × 10
-4the clenobuterol hydrochloride aqueous solution of mol/L;
B. 5 μ L α-methacrylic acids being joined 20mL concentration is 3 × 10
-4in mol/L clenobuterol hydrochloride aqueous solution, ultrasonic process 10 minutes, add 0.0028g N again, N-methylene-bisacrylamide, continue ultrasonic process 20 minutes, hold over night, obtains mixed liquor (mol ratio of described clenobuterol hydrochloride, α-methacrylic acid, N, N-methylene-bisacrylamide addition is 9.78:1:3.03);
C. be the ammonium persulfate solution of 0.5mol/L by 100 μ L concentration, slowly join in the mixed liquor that step 2 obtains, ultrasonic process 15 minutes initiated polymerizations, namely obtain clenobuterol hydrochloride molecularly imprinted polymer;
2. pair clenobuterol hydrochloride has the preparation of the nitrocellulose filter of Selective recognition ability: nitrocellulose filter is put into clenobuterol hydrochloride imprinted polymer solution and soak 30 minutes, take out rearmounted 40 DEG C of drying in oven, with 25% ethanol washing nitrocellulose filter, wash nitrocellulose filter with water again, dry, cut out;
3. cutting out size is 8cm × 1cm substrate, in substrate, cut out out a size is the void region of 4cm × 0.8cm, apply glue at the substratel of void region, be bonded in substrate by what cut out in above-mentioned steps 2 to the nitrocellulose filter that clenobuterol hydrochloride has a Selective recognition ability, cover the substrate that pressure one has identical void region again, to obtain final product.
Embodiment 2
A preparation method for molecular engram test strips, comprises the following steps:
1. the preparation of clenobuterol hydrochloride imprinted polymer solution:
A. compound concentration is 3 × 10
-5the clenobuterol hydrochloride aqueous solution of mol/L;
B. 3 μ L α-methacrylic acids being joined 10mL concentration is 3 × 10
-5in mol/L clenobuterol hydrochloride aqueous solution, ultrasonic process 5 minutes, add 0.0014g N again, N-methylene-bisacrylamide, continue ultrasonic process 10 minutes, hold over night, obtains mixed liquor (mol ratio of described clenobuterol hydrochloride, α-methacrylic acid, N, N-methylene-bisacrylamide addition is 117:1:30);
C. be the ammonium persulfate solution of 0.5mol/L by 80 μ L concentration, slowly join in the mixed liquor that step 2 obtains, ultrasonic process 10 minutes initiated polymerizations, namely obtain clenobuterol hydrochloride molecularly imprinted polymer;
2. pair clenobuterol hydrochloride has the preparation of the nitrocellulose filter of Selective recognition ability: nitrocellulose filter is put into clenobuterol hydrochloride imprinted polymer solution and soak 30 minutes, take out rearmounted 40 DEG C of drying in oven, with 15% ethanol washing nitrocellulose filter, wash nitrocellulose filter with water again, dry, cut out;
3. cutting out size is 8cm × 1cm substrate, in substrate, cut out out a size is the void region of 4cm × 0.8cm, apply glue at the substratel of void region, be bonded in substrate by what cut out in above-mentioned steps 2 to the nitrocellulose filter that clenobuterol hydrochloride has a Selective recognition ability, cover the substrate that pressure one has identical void region again, to obtain final product.
Embodiment 3
A preparation method for molecular engram test strips, comprises the following steps:
1. the preparation of clenobuterol hydrochloride imprinted polymer solution:
A. compound concentration is 3 × 10
-3the clenobuterol hydrochloride aqueous solution of mol/L;
B. 7 μ L α-methacrylic acids being joined 30mL concentration is 3 × 10
-3in mol/L clenobuterol hydrochloride aqueous solution, ultrasonic process 15 minutes, add 0.0042g N again, N-methylene-bisacrylamide, continue ultrasonic process 30 minutes, hold over night, obtains mixed liquor (mol ratio of described clenobuterol hydrochloride, α-methacrylic acid, N, N-methylene-bisacrylamide addition is 274:300:90);
C. be the ammonium persulfate solution of 0.5mol/L by 120 μ L concentration, slowly join in the mixed liquor that step 2 obtains, ultrasonic process 20 minutes initiated polymerizations, namely obtain clenobuterol hydrochloride molecularly imprinted polymer;
2. pair clenobuterol hydrochloride has the preparation of the nitrocellulose filter of Selective recognition ability: nitrocellulose filter is put into clenobuterol hydrochloride imprinted polymer solution and soak 30 minutes, take out rearmounted 40 DEG C of drying in oven, with 40% ethanol washing nitrocellulose filter, wash nitrocellulose filter with water again, dry, cut out;
3. cutting out size is 8cm × 1cm substrate, in substrate, cut out out a size is the void region of 4cm × 0.8cm, double faced adhesive tape is stained with at the substratel of void region, be bonded in substrate by what cut out in above-mentioned steps 2 to the nitrocellulose filter that clenobuterol hydrochloride has a Selective recognition ability, cover the substrate that pressure one has identical void region again, to obtain final product.
Two, kit
1. activity is the preparation of the glucose oxidase enzyme labeling clenobuterol hydrochloride solution of 200-400ug/mL
Take 5.0mg glucose oxidase to be dissolved in 1mL water, add the 0.1mol/L NaIO that 0.1 ~ 0.3mL newly joins
4solution, under room temperature, lucifuge stirs 10 ~ 30 minutes; Load in MW4000 bag filter by the solution be stirred, be the HAc-NaAc damping fluid dialysis of 4.4 to 1mmol/L pH value, 4 DEG C are spent the night; Get supernatant, adding 10 ~ 40 μ L 0.2mol/L pH value is the carbonate buffer solution of 9.5, makes the pH value of glucose oxidase be increased to 9.0 ~ 10.0, obtains glucose oxidase solution; Get 10mL color comparison tube, add 7mg clenobuterol hydrochloride and 1 ~ 3mL 0.01mol/L pH value is the carbonate buffer solution of 9.5, after it dissolves completely, add above-mentioned glucose oxidase solution, mixing, room temperature lucifuge stirs 1 ~ 3h, then adds the 4mg/mL NaBH that 0.1mL newly joins
4solution, mixing, places 1 ~ 3h, obtains mixed solution in the environment of 4 DEG C; Loaded by gained mixed solution in bag filter, be the phosphate buffer dialysis of 7.4 to 0.15mol/L pH value, 4 DEG C are spent the night; After having dialysed, taken out by the solution in bag filter, and keep in Dark Place in 4 DEG C, obtain glucose oxidase enzyme labeling clenobuterol hydrochloride solution, it is 200 ~ 400ug/mL that microplate reader measures its concentration.
2. standard color comparison card preparation
Example 1 method obtains clenobuterol hydrochloride molecular engram test strips 6, the clenobuterol hydrochloride solution of the above-mentioned glucose oxidase enzyme labeling prepared is dripped respectively in its reaction zone, incubation 10 minutes, after 40 DEG C of oven dry, with 70 μ L water wash reaction zones; In the reaction zone of drip washing, drip 40 μ L concentration is respectively 0 (i.e. blank solution), 3 × 10
-4mol/L, 3 × 10
-5mol/L, 3 × 10
-6mol/L, 3 × 10
-7mol/L, 3 × 10
-8the clenobuterol hydrochloride standard solution of mol/L, competitive reaction 10 minutes, then use 70 μ L water wash reaction zones; In the reaction zone of drip washing, drip 20 μ L concentration is respectively 0.1g/mL glucose solution, then to drip 20 μ L concentration be respectively 8 × 10
-5the red Y solution of administration of mol/L, enzymic catalytic reaction 5 minutes, by test strips 40 DEG C of oven dry, puts the colour developing situation of test strip under the uviol lamp of 365nm.
To detect that test strips is taken pictures under 365nm uviol lamp, see Fig. 2, according to the color of each test strips, make the standard color comparison card of 6 kinds of concentration, for detection testing sample time as a reference, convenient carries out quantitative and qualitative analysis.
3. kit
The embodiment 1 molecular engram test strips prepared above-mentioned, active glucose oxidase enzyme labeling clenobuterol hydrochloride solution, 0.1g/mL glucose solution, 3 × 10 for 200-400ug/mL
-4mol/L affixes one's name to red Y solution and standard color comparison card, puts into kit.During outdoor detection, then put into purple light pen, realize quick online detection.
Three, qualitative and semiquantitative detection method
Embodiment 4
The preparation of need testing solution: take 10g feed, adds 20mL 1mol/L dissolving with hydrochloric acid, then adds 180mL water, lucifuge shakes 25 minutes, with the speed of 3000r/min centrifugal 30 minutes, gets supernatant, regulate solution ph to be 8.0 by 1mol/L NaOH solution, to obtain final product.
Example 1 method obtains clenobuterol hydrochloride molecular engram test strips 1, the glucose oxidase enzyme labeling clenobuterol hydrochloride solution that the above-mentioned activity prepared is 200-400ug/mL is dripped respectively in its reaction zone, incubation 10 minutes, after 40 DEG C of oven dry, with 70 μ L water wash reaction zones; In the reaction zone of drip washing, drip the above-mentioned need testing solution prepared of 25 μ L respectively, competitive reaction 10 minutes, then use 70 μ L water wash reaction zones; In the reaction zone of drip washing, drip 20 μ L concentration is respectively 0.1g/mL glucose solution, then to drip 20 μ L concentration be respectively 8 × 10
-5the red Y solution of administration of mol/L, enzymic catalytic reaction 5 minutes, by test strips 40 DEG C of oven dry, puts the colour developing situation of test strip under the uviol lamp of 365nm, sees Fig. 3.
Result: because test strips fluorescence developing is included in standard color comparison card fluorescence color variation range, namely illustrates that this is quilitative method containing clenobuterol hydrochloride in this feed.Through conscientious and standard color comparison card comparison, because in test strips fluorescence color and standard color comparison card, the 5th color is close from left to right, therefore judge that the content order of magnitude that in feed need testing solution, the clenobuterol hydrochloride content order of magnitude and standard color comparison card the 5th color represents is identical, namely contains 10 in feed need testing solution
-7the clenobuterol hydrochloride of the mol/L order of magnitude, this is semi-quantitative detection method.
Claims (7)
1. a molecular engram test strips, comprises substrate and reaction zone, it is characterized in that: described reaction zone is nitrocellulose filter clenobuterol hydrochloride to Selective recognition ability covering substrate void region;
Described preparation method clenobuterol hydrochloride to the nitrocellulose filter of Selective recognition ability is: nitrocellulose filter is put into clenobuterol hydrochloride imprinted polymer solution and soak 30 minutes, take out rearmounted 40 DEG C of drying in oven, with 15-40% ethanol washing nitrocellulose filter, wash nitrocellulose filter with water again, dry, to obtain final product;
Described clenobuterol hydrochloride imprinted polymer solution take clenobuterol hydrochloride as template molecule, take α-methacrylic acid as function monomer, interact with hydrogen bond formation, add crosslinking chemical N, N-methylene-bisacrylamide, initiator ammonium persulfate occur free-radical polymerized, obtain clenobuterol hydrochloride imprinted polymer solution; The mol ratio of described template molecule, function monomer, crosslinking chemical is 117-274:1-300:30-90.
2. molecular engram test strips according to claim 1, is characterized in that: the material of described substrate is plastics or photo paper.
3. a preparation method for molecular engram test strips as claimed in claim 1 or 2, is characterized in that, comprise the following steps:
(1) preparation of clenobuterol hydrochloride imprinted polymer solution:
A. compound concentration is 3 × 10
-3-3 × 10
-5the clenobuterol hydrochloride aqueous solution of mol/L;
B. 3-7 μ L α-methacrylic acid being joined 10-30 mL concentration is 3 × 10
-3-3 × 10
-5in mol/L clenobuterol hydrochloride aqueous solution, ultrasonic process 5-15 minute, then add 0.0014-0.0042 g N, N-methylene-bisacrylamide, continue ultrasonic process 10-30 minute, hold over night, obtains mixed liquor;
C. be the ammonium persulfate solution of 0.5mol/L by 80-120 μ L concentration, slowly join in the mixed liquor that step b obtains, ultrasonic process 10-20 minute, namely obtains clenobuterol hydrochloride molecularly imprinted polymer;
(2) clenobuterol hydrochloride is had to the preparation of the nitrocellulose filter of Selective recognition ability: nitrocellulose filter is put into clenobuterol hydrochloride imprinted polymer solution and soak 30 minutes, take out rearmounted 40 DEG C of drying in oven, with 15-40% ethanol washing nitrocellulose filter, wash nitrocellulose filter with water again, dry, cut out;
(3) substrate is cut out, a void region is cut out out in substrate, apply glue at the substratel of void region or be stained with double faced adhesive tape, be bonded in substrate by what cut out in above-mentioned steps (2) to the nitrocellulose filter that clenobuterol hydrochloride has a Selective recognition ability, cover the substrate that pressure one has identical void region again, to obtain final product.
4. the preparation method of molecular engram test strips according to claim 3, is characterized in that, in described step (3), the size of substrate is 8 cm × 1cm, and the size of described void region is 4 cm × 0.8 cm.
5. the application of molecular engram test strips as claimed in claim 1 or 2, is characterized in that: for clenobuterol hydrochloride in testing sample qualitative, sxemiquantitative or quantitatively detect.
6. the kit containing, for example molecular engram test strips described in claim 1 or 2, it is characterized in that, comprise molecular engram test strips, active glucose oxidase enzyme labeling clenobuterol hydrochloride solution, 0.1g/mL glucose solution, 3 × 10 for 200-400ug/mL
-4mol/L affixes one's name to red Y solution and standard color comparison card.
7. the kit of molecular engram test strips according to claim 6, is characterized in that, also comprise purple light pen.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106501243A (en) * | 2016-10-01 | 2017-03-15 | 桂林理工大学 | A kind of method of use molecular engram test strips quick detection tripolycyanamide |
| CN110455755A (en) * | 2019-07-23 | 2019-11-15 | 西南交通大学 | The preparation method of quinolones sensor precursor and quinolones sensor |
| CN111961153A (en) * | 2020-08-26 | 2020-11-20 | 西南大学 | Thiodiglycol molecularly imprinted polymer and preparation method and application thereof |
| CN112114132A (en) * | 2020-08-28 | 2020-12-22 | 瑞捷生物科技江苏有限公司 | Carrier fixed with nitrocellulose membrane, preparation method and application thereof |
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| CN106501243B (en) * | 2016-10-01 | 2019-10-11 | 桂林理工大学 | A method of melamine is quickly detected with molecular engram test strips |
| CN110455755A (en) * | 2019-07-23 | 2019-11-15 | 西南交通大学 | The preparation method of quinolones sensor precursor and quinolones sensor |
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| CN111961153A (en) * | 2020-08-26 | 2020-11-20 | 西南大学 | Thiodiglycol molecularly imprinted polymer and preparation method and application thereof |
| CN111961153B (en) * | 2020-08-26 | 2022-03-18 | 西南大学 | Thiodiglycol molecularly imprinted polymer and preparation method and application thereof |
| CN112114132A (en) * | 2020-08-28 | 2020-12-22 | 瑞捷生物科技江苏有限公司 | Carrier fixed with nitrocellulose membrane, preparation method and application thereof |
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