Clenbuterol is a class of drugs, not a specific substance, which is a feed additive that promotes the growth of lean meat. Any substance that promotes the growth of lean meat and inhibits the growth of fat can be called "clenbuterol." Currently, substances that can do this are a class of drugs called beta-agonists, such as clenbuterol, which causes poisoning in China, and Ractopamine, which is allowed in the United States. . In China, what is commonly referred to as "clenbuterol" refers to Clenbuterol. It was once used as a medicine to treat bronchial asthma and was later banned due to its side effects.
Chinese name: Lean meat; Clenbuter; scientific name Clenbuterol; is an antiasthmatic drug. The drug is neither a veterinary drug nor a feed additive, but an adrenal nerve stimulant. Dichlorhydrin hydrochloride; ketoprofen; ammonia sulphate; ammonia succinimide; ammonia dichlorothreathine; ammonia chlorhydramine.
English name: CLENBUTEROL
CAS: 37148-27-9
Physical and chemical properties: white or white-like crystalline powder, odorless, bitter, melting point 161 ° C, soluble in water, ethanol, slightly soluble in acetone, insoluble in ether.
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Detection method
1. Gas chromatography-mass spectrometry (GC-MS)
The GC-MS method combines the efficient and rapid separation of chromatograms with the qualitative analysis of high-sensitivity mass spectrometry. It can qualitatively and quantitatively analyze a specific residue in the presence of multiple residues. Higher detection limits. Fente C. A et al. detected the residual CLB in bovine hair by GC-MS with a minimum detection limit of 5 ng/g. Pteer Batioens was applied to the tissues of cattle, sheep and pigs by gas chromatography-tandem mass spectrometry (GC-MS-MS). CLB content was tested with a minimum detection limit of 2 ng/g; Liu Qi et al. used GC-MS (EI ion source) to detect CLB in pig urine with a detection limit of 0.5 ng/mL; VanRhijin et al. used trimethylsilyl or 2-Dimethylsilylmorpholine derivatives detect CLB in urine extracts, and the derivatives are scanned by electrical pulse or chemical ionization to produce higher sensitivity. In addition, the GC-MS method has higher detection sensitivity and lower false positive rate than the HPLC method. Therefore, China has legalized GC-MS as a confirmatory method for detecting CLB (NY/T468~2001).
2. High performance liquid chromatography (HPLC)
HPLC is suitable for the determination of thermally unstable and highly polar β-agonists and their metabolites. Moreover, HPLC can be combined with pre-column extraction, purification, post-column fluorescence derivatization and mass spectrometry (MS) systems for easy analysis. Automation of the process. Huang Shixin et al (1995) applied UV detector to detect CLB residues in pig liver and pork at λ=243nm, column: shimpackCLC-ODS150×6.0mn, flow rate: 1mL/min, column temperature: room temperature 30°C. The minimum detection limit is 2 ng/g [4]. In foreign countries, HPLC (diode array detector) was used to determine CLB residues in animal foods. The minimum detection limit was 1.26 ng/g, and the recovery rate was 98.9% [5]. At present, HPLC has been used as a semi-positive method for detecting CLB residues in China. The minimum detection limit is 1-15 ng/g, which has the advantages of good specificity, strong selectivity, high detection accuracy, and low false positive rate. The disadvantage is that the sample processing time is long, the detection process is cumbersome, difficult to operate, and requires expensive instruments, which is subject to certain limitations in practical applications.
3. Enzyme-linked immunosorbent assay (ELISA)
The plant horseradish peroxidase (HRP) is chemically combined with clenbuterol (CL) to form an enzyme-coupled clenbuterol using immunological antigen-antibody specific binding and high-efficiency catalysis of the enzyme. The coated antibody (goat anti-rabbit IgG antibody) on the solid phase carrier is combined with a specific anti-Clenbuter antibody, and then the Clenbuterol and the enzyme-conjugated Clenbuter are added, which are competitive with The clenbuterol antibody binds, and after washing, a substrate is added, and the amount of clenbuterol to be measured is measured according to the change of the colored substance. If Clenbuterol is to be tested, the bound enzyme is less coupled with Clenbuterol, and the amount of colored matter is less. The Clenbuterol content in the sample is determined by visual or colorimetric method. The optimal wavelength for colorimetric is 450 nm and the reference wavelength should be greater than 600 nm.
4, Colloidal gold immunochromatography (Colloidal gold immunochromatography)
Using competitive colloidal gold immunochromatography, Clen in the test solution binds to the gold-labeled antibody on the gold-labeled pad to form a complex. If the concentration of Clen in the test solution is lower than the sensitivity value, the unbound gold-labeled antibody flows to the T-region. When the Clen-BSA conjugate immobilized on the membrane is combined, it gradually aggregates into a visible T line; if the Clen concentration is higher than the sensitivity value, the gold standard antibody forms a complex and will not be Clen- with the T line. The BSA conjugates combine to form a visible T line. The unfixed composite stream passes through the T zone and is captured by the secondary antibody of the C zone and forms a visible C line. The appearance of the C line indicates that immunochromatography occurs, that is, the test paper is effective.
Testing method
1. Read the instruction manual thoroughly before performing the test. Return the reagent plate and the sample solution to be tested to room temperature before use.
2. Peel the aluminum plastic bag and remove the test paper.
3. Operate according to the model (urine should not directly wet the observation area).
3.1 Dip the white end of the test paper into the urine (the liquid level must not exceed the horizontal line) for 5 seconds;
3.2 Pipette the urine of the sample to be tested with a dropper, and slowly add 3 drops of the sample to the well.
4. Place the test strip flat for 1 minute and wait for the red strip to appear.
5. Read the results within 3 to 8 minutes, and the interpretation is invalid after 10 minutes.
6. [1] Results can be detected within 10 minutes
Result interpretation
Positive ( ): Only a purple-red band appears in the quality control area (C). No purple-red bands appeared in the test area (T).
Negative (-): Two purple-red bands appear. One is located in the test area (T) and the other is located in the quality control area (C).
Invalid: The purple control strip does not appear in the quality control area (C), indicating that the test failed or the test strip has failed.
Note: The fuchsia strips in the test area (T) show a color shade. However, during the specified observation time, regardless of the color depth of the ribbon, even a very weak ribbon should be judged as a negative result.
Precautions
1. Please use the test card once during the warranty period.
2. Avoid direct sunlight and direct blow of the electric fan during testing.
3. Try not to touch the white film surface in the center of the test card.
4. Urine sample dropper should not be mixed to avoid cross contamination.
5. If the urine sample shows sediment or turbidity, please centrifuge and test again.
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