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Sample pretreatment method for detecting different clenbuterol residuals

A sample pretreatment and clenbuterol technology, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of regulatory vacuum, large influence of reagent environment, low precision and accuracy, and achieve the effect of expanding the scope

Inactive Publication Date: 2014-01-15
CHINA ANIMAL HEALTH & EPIDEMIOLOGY CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The current analysis methods have problems such as cumbersome detection process, large impact on the environment of the reagents used, and high technical requirements for operation.
In actual application, testing institutions are prone to unstable test results, low precision and accuracy, which can easily lead to misjudgment of test results, missed tests or false positives, which lead to the administrative department's "clenbuterol" "lack of regulation
[0007] More importantly, at present, there is no effective method to simultaneously detect these "clenbuterol" drugs prohibited by the Ministry of Agriculture at home and abroad. There is no effective technical means for the supervision of the illegal use of "Clenbuterol", which will easily cause a "regulatory vacuum"

Method used

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  • Sample pretreatment method for detecting different clenbuterol residuals
  • Sample pretreatment method for detecting different clenbuterol residuals

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Accurately weigh 2 g of blood, body fluid and other liquid samples or muscle tissue into a 50 mL centrifuge tube, add 5 mL of ammonium acetate solution (0.2 mol / L) at pH 5.2, and then add 20 μL of glucuronidase-sulfatase (sigma company) Vortex for 30s, place at 37°C overnight, take it out the next day, 4 Centrifuge at 000rpm for 5min, take the supernatant and put it in another centrifuge tube; add 5mL of 1% methanol solution of formic acid to the residue, vortex for 30s, centrifuge at 4000rpm for 5min, combine the supernatant, then add 5mL of 1% methanol solution of formic acid to extract once, and combine the supernatant liquid, to be purified.

Embodiment 2

[0015] Accurately weigh 2 g of liver, kidney, lung and other tissue samples into a 50 mL centrifuge tube, except adding 5 mL of 1% formic acid methanol solution, vortexing for 30 s, and centrifuging at 10,000 rpm for 5 min, other steps are the same as in Example 1.

Embodiment 3

[0017] Take 5mL of the extracted solution and add it to the MCX solid-phase extraction column activated by 3mL methanol and pure water for purification, wash with 3mL pure water and methanol in turn, drain the MCX purified extraction column, elute with 3mL of 5% ammoniated methanol solution, blow with nitrogen After drying, dissolve it with 0.1% formic acid aqueous solution / acetonitrile (9:1, v / v), and obtain satisfactory results after detection by liquid chromatography-tandem mass spectrometer (quantification by internal standard method). The specific detection parameters are as follows:

[0018] 1. Sensitivity of the method: the detection limit of 21 drugs in blood and muscle can reach 0.1 μg / kg, and the limit of quantification is 0.25 μg / kg; the detection limit of 21 drugs in liver, kidney and lung tissue can reach 0.2 μg / kg, The limit of quantification is 0.5 μg / kg, which is greatly improved or equivalent compared with the existing detection method, and can fully meet the ...

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PUM

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Abstract

The invention relates to a sample pretreatment method for detecting different clenbuterol residuals. In the method, no halogen preparation is used, liquid-liquid extraction processes after alkalization are reduced, and the operation process is simple and fast. The method adopts an enzymolysis, extraction and purification manner for sample pretreatment, particularly, a zymolytic sample is directly purified through an MCX solid phase extraction column after being extracted through acidified methyl alcohol, and detection is performed through liquid chromatography tandem mass spectrometry. Twenty four samples can be detected within 6 hours, so that the detecting time is saved by about half, and the detecting cost is also lowered greatly.

Description

Technical field: [0001] The invention relates to a biological detection method, in particular to a sample pretreatment method for detecting different types of "clenbuterol" residues. Background technique: [0002] "Clenbuterol", originally a common name for clenbuterol hydrochloride, now generally refers to a class of substances containing β-adrenergic receptor agonists illegally used by the farming industry to promote animal growth and increase lean meat percentage. Many countries, including my country, have regulated β in food animals 2 - The illegal use of receptor agonists has been strictly supervised. Since 2002, my country has issued Announcements No. 176, 193, and 235. In 2010, Announcement No. 1519 was issued, clearly prohibiting the use of Clenium hydrochloride Terol, albuterol, salbutamol sulfate, ractopamine, dopamine hydrochloride, cimaterol, terbutaline sulfate, phenylethanolamine A, bambuterol, zilpaterol hydrochloride, chlorprenaline hydrochloride, mabuterol ,...

Claims

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Application Information

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IPC IPC(8): G01N30/06
Inventor 赵思俊王君玮曲志娜王娟孙晓亮曹旭敏王玉东洪军
Owner CHINA ANIMAL HEALTH & EPIDEMIOLOGY CENT
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