CN215953631U - Clenbuterol hydrochloride short-term test card - Google Patents

Abstract

The utility model discloses a clenbuterol hydrochloride rapid detection card which comprises a box body (1), wherein the box body (1) comprises a dropper storage area (2), a quality control product storage area (3), a specification storage area (4) and a clenbuterol hydrochloride rapid detection card storage area (5). The quality control product is added in the clenbuterol hydrochloride detection card, so that on one hand, real-time quality control is provided for the detection of clenbuterol hydrochloride, on the other hand, the detection of a customer when an abnormal result occurs is facilitated, so that whether the sample or the operation is a problem or the problem of the detection card is determined, and the customer can conveniently verify the abnormal condition. The clenbuterol hydrochloride rapid detection card provided by the utility model is suitable for detecting clenbuterol hydrochloride in animal urine and the like, has the advantages of strong specificity, high sensitivity, good stability and high detection speed, and can be used for screening clenbuterol hydrochloride.

Description

Clenbuterol hydrochloride short-term test card

Technical Field

The utility model relates to the technical field of immunochemistry rapid detection, in particular to a clenbuterol hydrochloride detection card.

Background

Clenbuterol is a general name of a class of medicines, and any substance capable of inhibiting the generation of animal fat and promoting the growth of the clenbuterol is called as 'clenbuterol'. Substances capable of achieving such functions are mainly drugs called beta-receptor agonists (also called beta-agonists), among which clenbuterol hydrochloride, salbutamol, ractopamine and the like are more common.

After the clenbuterol hydrochloride enters the livestock body, the metabolic pathway of nutrients can be changed, the growth of animal muscles is promoted, particularly the synthesis of skeletal muscle protein is promoted, the conversion and decomposition of fat are accelerated, and the lean meat percentage of the livestock is improved. Therefore, in order to promote the growth of animals and improve the feed conversion rate and the lean meat percentage of animals, some illegal vendors use the clenbuterol hydrochloride as a feed additive, so that the content of the clenbuterol hydrochloride in the animal bodies exceeds the standard. And if people eat livestock meat with the clenbuterol hydrochloride content exceeding the standard for a long time, adverse reactions occur to the body, and even the life is threatened in serious cases. Therefore, people need to improve vigilance and strictly monitor the clenbuterol hydrochloride content in the livestock.

Currently, there are various methods for detecting clenbuterol hydrochloride, for example: gas chromatography-mass spectrometry, high performance liquid chromatography, enzyme-linked immunosorbent assay, and colloidal gold assay. Among them, the colloidal gold method is the most popular method for clenbuterol hydrochloride detection because of its advantages such as simple detection process and low cost. However, there are cases where the C-line is not established (due to a production accident or the like) and quality control is poor in the colloidal gold assay. Therefore, there is an urgent need for a new type of gold colloid that can ensure the establishment of the C-line (reduce the invalid detection rate) and the built-in quality control.

SUMMERY OF THE UTILITY MODEL

In order to make up the defects of the prior art, the utility model aims to provide a clenbuterol hydrochloride rapid detection card.

Therefore, the utility model provides a clenbuterol hydrochloride rapid detection card, which comprises a box body (1), wherein the box body (1) comprises a dropper storage area (2), a quality control product storage area (3), an instruction book storage area (4) and a clenbuterol hydrochloride rapid detection card storage area (5).

Preferably, 50 bags of clenbuterol hydrochloride rapid detection cards are stored in the clenbuterol hydrochloride rapid detection card storage area (5); each clenbuterol hydrochloride rapid detection card comprises a PVC plate, and a sample pad, a gold label pad, a nitrocellulose membrane and a water absorption pad are sequentially fixed on the PVC plate; the gold-labeled pad is colloidal gold labeled clenbuterol hydrochloride antibody; the surface of the nitrocellulose membrane is marked with a detection line and a quality control line, wherein the quality control line is a mixed solution of rabbit anti-mouse IgG and active blue, and the detection line is clenbuterol hydrochloride antigen.

Preferably, one quality control material is stored in the quality control material storage area (3) of the utility model, and the volume of the quality control material is 0.5 ml/piece; the quality control product is clenbuterol hydrochloride antigen.

Preferably, the concentration of the clenbuterol hydrochloride antigen in the utility model is 20 ppb.

Preferably, 50 droppers are stored in the dropper storage area (2) of the utility model.

The quality control product is added in the clenbuterol hydrochloride detection card, so that on one hand, real-time quality control is provided for the detection of clenbuterol hydrochloride, on the other hand, the detection of a customer when an abnormal result occurs is facilitated, so that whether the sample or the operation is a problem or the problem of the detection card is determined, and the customer can conveniently verify the abnormal condition.

According to the utility model, the active blue is added at the position of the C line, so that the C line is more uniform and more stable, meanwhile, the T line is not interfered, and the possibility of false positive caused by C gold is reduced. When the sample runs to the C line, the blue color of the C line changes into red (the active blue moves upwards along with the liquid, and the gold-labeled antibody is combined with the rabbit anti-mouse IgG to form a red strip), and whether the detection card fails or not is further detected. In addition, whether production is normal or not, whether production accidents occur or not and the like can be conveniently judged in the production process of the detection card.

The clenbuterol hydrochloride rapid detection card provided by the utility model is suitable for detecting clenbuterol hydrochloride in animal urine and the like, has the advantages of strong specificity, high sensitivity, good stability and high detection speed, and can be used for screening clenbuterol hydrochloride.

Drawings

FIG. 1 shows the assembly of a fast clenbuterol hydrochloride test card (large panel).

Fig. 2 is a diagram of a finished product of an inner package of the clenbuterol hydrochloride rapid detection card, wherein a line C is a blue strip.

Fig. 3 is a schematic diagram of a packaged finished product of the clenbuterol hydrochloride rapid detection card, wherein 1 is a box body, 2 is a dropper storage area, 3 is a quality control product storage area, 4 is an instruction book storage area, and 5 is a clenbuterol hydrochloride rapid detection card storage area.

FIG. 4 is a diagram showing the detection judgment of the fast clenbuterol hydrochloride card, wherein the line C and the line T are red strips in the case of negative judgment; when positive judgment is carried out, the line C is a red strip, and the line T has no strip; when the judgment is invalid, the line C is a blue stripe or no stripe, and the line T is a red stripe or no stripe.

Detailed Description

The following detailed description of the preferred embodiments of the present invention, taken in conjunction with the accompanying drawings, will make the advantages and features of the utility model easier to understand by those skilled in the art, and thus will clearly and clearly define the scope of the utility model. The experimental procedures not described in detail in the examples are generally carried out according to the routine procedures in the art or according to the conditions recommended by the manufacturers. The reagents and drugs mentioned in the examples are all common commercial products unless otherwise specified.

EXAMPLE 1 preparation of fast detection card for clenbuterol hydrochloride

1 the PVC base plate used as a manufacturing material is purchased from Hangzhou mountain navigation science and technology limited company; the nitrocellulose membrane is purchased from Shantou and biological medicine technology (Shantou) Co.Ltd; the glass cellulose membrane is purchased from Hangzhou Huarun paper industry technology development company; the absorbent paper is purchased from Hangzhou mountain navigation science and technology limited; the clenbuterol hydrochloride antigen is purchased from the center of the Luoyang Bai' otong experimental material; the clenbuterol hydrochloride antibody is purchased from the center of the Luoyang Bai' otong experimental material; active blue was purchased from Shanghai Ji to Biochemical technology, Inc.; rabbit anti-mouse IgG (1mg/ml) was purchased from Biotech, Inc. of Solebao, Beijing.

2 preparation of nitrocellulose membrane and quality control product

2.1 preparation of coating buffer 1g of sucrose was added to 100mL of PBS (0.01mol/L, pH 7.2) to prepare a coating buffer, which was sterilized by filtration through a 0.22 μm filter and stored at 2-8 ℃ for further use.

2.2 preparation of quality control line (line C) solution active blue is added into ultrapure water to prepare 1mol/L active blue solution for standby. The active blue solution and rabbit anti-mouse IgG 1:9 are mixed evenly for standby.

2.3 preparation of the nitrocellulose membrane, adhering the nitrocellulose membrane to a corresponding position of a PVC (polyvinyl chloride) base plate, diluting a clenbuterol hydrochloride antigen to 1mg/mL by using a coating buffer solution, adjusting the scribing position and height of a scribing machine, scribing to form a T line, namely a detection line, wherein the T line is close to a gold mark pad end, adjusting the scribing position and height of the scribing machine by using a quality control line (C line) solution, scribing to form a C line, namely a quality control line, wherein the C line is close to an absorption pad, and the distance between the two lines is 5-8 mm. Drying in a 37 ℃ oven for 15 hours, sealing in an aluminum foil bag filled with a drying agent, and storing at 2-30 ℃ for later use.

2.4 preparation of quality control product clenbuterol hydrochloride antigen is diluted by using coating buffer solution, the concentration of the diluted clenbuterol hydrochloride antigen is 20ppb, the clenbuterol hydrochloride antigen is quantitatively subpackaged, and each clenbuterol antigen is 0.5ml and is stored for later use at 2-8 ℃.

Preparation of 3 gold-labeled pad

3.1 preparing 1% chloroauric acid solution with colloidal gold, heating, stirring, adding 1% trisodium citrate solution, and boiling. Cooling and then standby. To 1mL of gold water, 5. mu.g of a clenbuterol hydrochloride antibody was added and reacted for 10 minutes, and then a 1% blocking agent (1% BSA in PBS) was added and blocked for 20 minutes. Then centrifuging and removing supernatant, adding the complex solution, and shaking uniformly.

3.2 spraying gold, namely spraying the resuspended colloidal gold solution on a glass cellulose membrane by using a gold spraying machine according to the amount of 2 mu L/cm, drying for 16-20 hours at 37 ℃, packaging by using an aluminum foil bag, and storing for later use at 2-30 ℃.

4 treatment of the sample pad

4.1 preparation of sample pad treating solution Casein, sodium cholate, surfactant S9, vinylpyrrolidone (PVP), Tris and ProClin 300 are weighed and added into ultrapure water to be dissolved, after being completely dissolved, the mixture is mixed evenly, the pH value is adjusted, ultrapure water is added to be constant volume, after being mixed evenly, the mixture is sterilized by a 0.22 mu m microporous filter membrane, and the mixture is stored for standby use at the temperature of 2-8 ℃.

4.2 evenly coating the sample pad treatment solution on a glass cellulose membrane, drying for 16-20 hours at 37 ℃, packaging by an aluminum foil bag, and storing for later use at 2-30 ℃.

5 clenbuterol hydrochloride rapid detection card assembly as shown in figure 1, cut sample pad, gold mark pad, water absorption pad into corresponding width and paste in proper order to the corresponding position of the PVC bottom plate that has been stuck with nitrocellulose membrane, make gold mark pad, water absorption pad contact with nitrocellulose membrane part separately, sample pad contacts with gold mark pad part, make the large plate.

6 packaging, cutting the large plate into test strips with the width of 3mm by using a slitter, installing a shell (figure 2), sealing and packaging by using an aluminum foil bag, containing one test strip and one suction tube, packing one desiccant, storing at 2-30 ℃ in a dark place for later use, and temporarily setting the storage period to be 24 months.

7 Assembly of test cards Each test card component that was qualified was assembled as follows (FIG. 3)

Components	Number of	Specification of
			Fast clenbuterol hydrochloride detecting card	50 bags	1 group/bag
Dropper	50 pieces	1 piece/bag
			Quality control product	1 count	0.5 ml/piece
Description	1 part of	/

8 test card usage and determination

8.1 methods of use

8.1.1 complete reading of the instructions before testing, the test card and the sample solution to be tested were returned to room temperature before use.

8.1.2 remove the test card from the original package and use it as soon as possible within an hour after opening.

8.1.3 the test card is placed horizontally, a dropper is used for absorbing the sample solution to be tested (generally urine, tissue fluid), 2-3 drops are vertically dropped into the sample adding hole, and timing is started after sample adding.

8.1.4 results should be read in 3-5 minutes, and the results are judged according to the schematic diagram when the interpretation is invalid at other times.

8.2 decision (FIG. 4)

8.2.1 negative (-): two red bands appear indicating that the sample does not contain clenbuterol hydrochloride or its concentration is below the limit of detection.

8.2.2 Positive (+): and detecting that the T line has no band, and the quality control C line is a red band, which indicates that the concentration of the clenbuterol hydrochloride in the sample is higher than the detection limit.

8.2.3 are invalid: the quality control C line is a blue strip all the time and is not changed into a red strip; or the quality control C-line has no strip, indicating an incorrect procedure or a failed test card.

8.2.4 when the detection is ambiguous, the quality control product can be used for correction, and if the detection result of the quality control product is negative, the batch of detection cards are invalid and cannot be used.

9 test card use notes

9.1 the detection card is required to be used once in the guarantee period;

9.2, the direct sunlight and the direct blowing of the electric fan are avoided during detection;

9.3 touch the white film surface in the center of the detection card is avoided as much as possible;

9.4 the sampling dropper can not be used in a mixed way so as to avoid cross contamination;

9.5 if the sample to be detected has sediment or turbid substances, please detect after centrifuging;

9.6 any problems encountered with the trial, please contact the supplier;

9.7 if the standard substance needs to be directly detected, please use the mixed negative pig urine for preparation.

10 storage and effective period of the detection card, namely, the detection card except the quality control product is stored in a shady and dry place at 2-30 ℃, and the effective period is 24 months; and the quality control product is stored in a shady and dry place at the temperature of 2-8 ℃, and the validity period is 24 months.

Example 2 evaluation of the Performance of the fast detection card of clenbuterol hydrochloride

1 reagent strip for preparing the clenbuterol hydrochloride rapid detection card, refer to the preparation of example 1, and the batch numbers are 210501, 210502 and 210505 respectively.

2, the prepared 3 batches of detection cards are used for detecting ractopamine quality control products, salbutamol quality control products and 10 negative samples, and the result shows that all the samples are negative.

TABLE 1 results of specific detection

Note: "+" indicates a positive test result, and "-" indicates a negative test result.

3-sensitivity test the detection results of 3 batches of detection cards on clenbuterol hydrochloride quality control products with different concentrations are shown in table 2, and the lowest detection limit is 3 ppb. Indicating that the sensitivity of the test card is better.

TABLE 2 results of sensitivity measurements

Note: "+" indicates a positive test result, and "-" indicates a negative test result.

4 repeated evaluation three batches of test cards were tested for strong positive, weak positive and negative samples according to the instructions, and the results are shown in table 3. As can be seen from the table, the 3 batches of the detection cards are consistent in batch and between batches for detecting the strong positive samples, the weak positive samples and the negative samples, which indicates that the clenbuterol hydrochloride rapid detection card prepared by a laboratory method has better repeatability.

TABLE 3 results of repeated measurements

The above description is only an embodiment of the present invention, and not intended to limit the scope of the present invention, and all modifications of equivalent structures and equivalent processes performed by the present specification and drawings, or directly or indirectly applied to other related technical fields, are included in the scope of the present invention.

Claims (3)

1. The clenbuterol hydrochloride rapid detection card is characterized by comprising a box body (1), wherein the box body (1) comprises a dropper storage area (2), a quality control product storage area (3), an instruction book storage area (4) and a clenbuterol hydrochloride rapid detection card storage area (5); the clenbuterol hydrochloride rapid detection card storage area (5) is used for storing 50 bags of clenbuterol hydrochloride rapid detection cards; each clenbuterol hydrochloride rapid detection card comprises a PVC plate, and a sample pad, a gold label pad, a nitrocellulose membrane and a water absorption pad are sequentially fixed on the PVC plate; the gold-labeled pad is colloidal gold labeled clenbuterol hydrochloride antibody; a detection line and a quality control line are marked on the surface of the nitrocellulose membrane, wherein the quality control line is a mixed solution of rabbit anti-mouse IgG and active blue, a blue strip is arranged on a detection card, and the detection line is a clenbuterol hydrochloride antigen; one quality control product is stored in the quality control product storage area (3), and the quantity of the quality control product is 0.5 ml/piece; the quality control product is clenbuterol hydrochloride antigen.

2. The detection card of claim 1, wherein the concentration of the clenbuterol hydrochloride antigen is 20 ppb.

3. The test card of claim 1, wherein 50 pipettes are stored in the pipette storage area (2).

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