Method for detecting Clenbuterol residual quantity in hair

A residue and hair technology, applied in measurement devices, instruments, scientific instruments, etc., can solve the problems of low detection sensitivity and specificity, limit clenbuterol residues, and complicated sample processing procedures, and achieve easy sampling and shortening of time. , Accurate and reliable qualitative and quantitative detection effect

Inactive Publication Date: 2012-01-04
CENT HOSPITAL XUHUI DISTRICT SHANGHAI CITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] Most of the existing clenbuterol detection technologies have shortcomings such as inconvenient sampling, complicated sample processing process, high sample processing cost, low detection sensitivity and specificity, which limit the application of clenbuterol residue detection in my country's food safety supply guarantee system. Therefore, it is of great practical significance to study the selection of convenient and reliable inspection materials, simple and fast sample pretreatment methods, and accurate and effective detection methods.

Method used

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  • Method for detecting Clenbuterol residual quantity in hair
  • Method for detecting Clenbuterol residual quantity in hair
  • Method for detecting Clenbuterol residual quantity in hair

Examples

Experimental program
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Effect test

Embodiment 1

[0033] Embodiment 1 pig hair sample pretreatment method

[0034] One case of negative blank pig hair and three cases of positive pig hair collected from pigs suspected to have been fed clenbuterol in a slaughterhouse, the hair samples were washed with water and methanol respectively, dried at room temperature, and then shredded to 2- 3mm fragments. Take 50mg of hair into a centrifuge tube, add 50μl of 25ng / ml internal standard working solution and 0.5ml of 2mol / L sodium hydroxide solution, vortex and mix for 1min, heat in a water bath at 90°C for 10min, take it out, and add excess Na 2 SO 4Solid, mix well, add 2.5ml ether, vortex for 5min, centrifuge at 15,000×g for 10min; absorb the upper organic phase, add 100μl of 2% (v / v) formic acid solution, vortex for 5min, centrifuge at 15,000×g for 10min; absorb the lower layer of water phase, transferred to a vial for injection.

Embodiment 2

[0035] Another pretreatment method of the pig hair sample of embodiment 2

[0036] In addition, 1 case of negative blank pig hair and 3 cases of positive pig hair in Example 1 were taken, washed with water and dichloromethane respectively, dried at room temperature, and then cut into fragments of 2-3 mm. Take 50mg of hair into a centrifuge tube, add 50μl of 25ng / ml internal standard working solution, and 1ml of 1mol / L potassium hydroxide solution, vortex and mix for 1min, heat in a water bath at 65°C for 15min, take it out, add excess NaCl solid, and mix well , add 2ml tert-butyl methyl ether, vortex for 5min, centrifuge at 15,000×g for 10min; absorb the upper organic phase, add 100μl 5% (v / v) acetic acid solution, vortex for 5min, centrifuge at 15,000×g for 10min; absorb the lower aqueous phase, transfer To the vial for injection.

Embodiment 3

[0037] Another pretreatment method of the pig hair sample of embodiment 3

[0038] Another negative blank and 3 positive pig hairs in Example 1 were taken, washed with water and 0.01mol / L dilute hydrochloric acid respectively, dried at room temperature, and then cut into fragments of 2-3 mm. Take 50mg of hair into a centrifuge tube, add 50μl of 25ng / ml internal standard working solution, 1ml of concentrated ammonia solution (25-28%, v / v), vortex and mix for 1min, heat in a water bath at 90°C for 15min, take it out, and add excess Na 2 SO 4 Solid, mix well, add 2ml of dichloromethane, vortex for 5min, centrifuge at 15,000×g for 10min; absorb the upper organic phase, add 200μl 0.5% (v / v) formic acid solution, vortex for 5min, centrifuge at 15,000×g for 10min; absorb the lower layer The aqueous phase was transferred to a vial for injection.

[0039] For the samples processed by the three pig hair sample processing methods in Examples 1, 2, and 3, the clenbuterol residue in the ...

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Abstract

The invention discloses a method for detecting clenbuterol residue in hair. The method sequentially comprises the following steps: (1) pre-processing a hair sample: hydrolyzing the sample with alkali at a high temperature, extracting the sample with an organic solvent and reversely extracting the sample with acid water; and (2) taking deuterated clenbuterol-D9 aqueous solution with the concentration of 5-500ng / ml as an internal standard, and detecting the clenbuterol residue in hair by liquid chromatogramy-tandem mass spectrometry. The method has the following advantages: 1. hair is taken as a detection material, which provides easy sampling, convenient storage and repeatable sampling, sampling is simple and rapid and has trace amount and high efficiency; 2. the pre-processing method of the hair sample is optimized, which shortens the pre-processing time, simplifies the processing steps and obviously reduces the cost; and 3. an isotopic compound is taken as the internal standard, and the liquid chromatogramy-tandem mass spectrometry is used as a detection means, the minimum detection limit reaches 0.01ng / g, and the minimum quantification limit is 0.5ng / g. The method has the accurate and reliable capacity to qualitatively and quantitatively detect the clenbuterol, and can be taken as a tool for large sample screening or small sample confirmation.

Description

technical field [0001] The invention relates to a method for detecting residual clenbuterol in hair, in particular to a method for quantitatively using deuterated clenbuterol as an internal standard by liquid chromatography-tandem mass spectrometry (LC-MS / MS method for short). Method for detection of clenbuterol residues in hair. Background technique [0002] Clenbuterol can significantly promote the growth of animals, improve fat distribution in animals and increase lean meat percentage, so it is commonly known as "clenbuterol". Based on the continuous occurrence of clenbuterol poisoning incidents caused by clinical consumption of animal viscera, the addition of clenbuterol drugs to feed is generally prohibited in countries all over the world. Since 1997, my country has repeatedly banned the use of clenbuterol in the animal husbandry industry, but poisoning incidents still occur frequently in various places, indicating that the phenomenon of illegal use of clenbuterol stil...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/06G01N30/02G01N30/34
Inventor 刘罡一张梦琪陆川李水军贾晶莹余琛
Owner CENT HOSPITAL XUHUI DISTRICT SHANGHAI CITY
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