Method for rapidly detecting beta receptor agonist in pork

A β-receptor and agonist technology, applied in the field of analysis and detection, can solve the problems of cumbersome process, large loss of repeated extraction, unsuitable for the detection of large batches of samples, etc., and achieve the effect of simple operation and shortened enzymatic hydrolysis time

Inactive Publication Date: 2019-02-01
ANHUI GUOKE TESTING TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the detection methods of β-receptor agonists include enzyme-linked immunoassay, gas chromatography-mass spectrometry, liquid chromatography-mass spectrometry, etc., but this method is lacking in detection efficiency and cost. The disadvantage of this kind of pretreatment process is that the loss of repeated extraction is large , the processing time is long, the economic cost is high, and the process is cumbersome, which is not suitable for the detection of large quantities of samples. Therefore, it is very necessary to establish a simple, fast and efficient detection method for β-receptor agonists in pork

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] 1) Weigh 2g of pork sample, add 8g of 0.2mol / L ammonium acetate buffer solution with a pH value of 5.0 to the pork sample, homogenize at 5000r / min for 1min, add 5000U of β-glucose hydrochloride to the homogenized pork sample Aldosidase and arylsulfatase, 35°C, enzymatic hydrolysis for 3 hours, add 20g of a mixture of isopropanol and ethyl acetate with a volume ratio of 7:5, vortex for 10 minutes, and centrifuge at 8000rpm for 5 minutes to obtain the supernatant, which is set aside ;

[0023] 2) Equilibrate the MCX solid-phase extraction column with 10ml of methanol, 10ml of deionized water, and 10ml of 0.1mol / L hydrochloric acid solution in sequence. After pretreatment of the MCX solid-phase extraction column, load the supernatant from step 1) onto the MCX The solid-phase extraction column was rinsed with deionized water and vacuum-drained the MCX solid-phase extraction column, and the MCX solid-phase extraction column was eluted with a mixture of ammonia and methanol a...

Embodiment 2

[0027] 1) Weigh 2g of pork sample, add 8g of 0.2mol / L ammonium acetate buffer solution with a pH value of 5.0 to the pork sample, homogenize at 5000r / min for 1min, add 5500U of β-glucose hydrochloride to the homogenized pork sample Alsidase and arylsulfatase, 37°C, enzymatic hydrolysis for 4 hours, add 20g of a mixture of isopropanol and ethyl acetate with a volume ratio of 7:5, vortex for 13 minutes, and centrifuge at 8000rpm for 7 minutes to obtain the supernatant, which is set aside ;

[0028] 2) Equilibrate the MCX solid-phase extraction column with 10ml of methanol, 10ml of deionized water, and 10ml of 0.1mol / L hydrochloric acid solution in sequence. After pretreatment of the MCX solid-phase extraction column, load the supernatant from step 1) onto the MCX The solid-phase extraction column was rinsed with deionized water and vacuum-drained the MCX solid-phase extraction column, and the MCX solid-phase extraction column was eluted with a mixture of ammonia and methanol at ...

Embodiment 3

[0032] 1) Weigh 2g of pork sample, add 8g of 0.2mol / L ammonium acetate buffer solution with a pH value of 5.0 to the pork sample, homogenize at 5000r / min for 1min, add 6000U of β-glucose hydrochloride to the homogenized pork sample Aldosidase and arylsulfatase, 39°C, enzymatic hydrolysis for 5h, add 20g of a mixture of isopropanol and ethyl acetate with a volume ratio of 7:5, vortex for 15min, and centrifuge at 8000rpm for 10min to obtain the supernatant, which is set aside ;

[0033] 2) Equilibrate the MCX solid-phase extraction column with 10ml of methanol, 10ml of deionized water, and 10ml of 0.1mol / L hydrochloric acid solution in sequence. After pretreatment of the MCX solid-phase extraction column, load the supernatant from step 1) onto the MCX The solid-phase extraction column was rinsed with deionized water and vacuum-drained the MCX solid-phase extraction column, and the MCX solid-phase extraction column was eluted with a mixture of ammonia and methanol at a volume rat...

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Abstract

The invention relates to a method for rapidly detecting beta receptor agonist in pork. The method comprises the steps of: 1) weighing a pork sample, carrying out homogenizing and enzymatic hydrolysis,adding an extracting solution, carrying out vortex oscillation for 10-15 minutes and centrifuging for 5-10 minutes at the speed of 8000rpm to obtain supernatant for later use; 2) After pretreating anMCX (Mixed-mode Cation eXchange) solid phase extraction column, loading the supernatant of the step 1) onto the MCX solid phase extraction column, washing by using deionized water, draining the MCX solid phase under negative pressure, eluting the MCX solid phase extraction column by using eluent, concentrating, drying and diluting by using a 0.1% of formic acid solution to obtain a sample liquid;and 3) loading the sample liquid onto an ultra-high performance liquid chromatograph to detect the content of the beta receptor agonist in the sample liquid. The method disclosed by the invention hasthe advantages that the enzymatic hydrolysis time is significantly shortened by optimizing the enzymatic hydrolysis conditions; the pork sample can be quickly pretreated through extraction and elution; a satisfactory separation effect can be quickly obtained when the sample loading and detection are carried out by using the ultra-high performance liquid chromatograph of the method; and the methodis simple and convenient to operate, fast and high in efficiency and is suitable for the detection of large-batch samples.

Description

technical field [0001] The invention relates to the field of analysis and detection, in particular to a method for rapid detection of beta receptor agonists in pork. Background technique [0002] β-receptor agonists are a class of chemically synthesized phenylethanolamines. Representative compounds are tobuterol, pumbuterol, clenbuterol, fenoterol, and cimaterol. It is used to prevent and treat bronchial asthma and bronchospasm in humans and animals. Later, it was found that adding this type of drug in the feed has a nutrient redistribution effect and can significantly increase the lean meat rate of animals. Therefore, it has been widely concerned as a growth-promoting additive. However, people will experience adverse reactions such as flushing, headache, dizziness, chest tightness, heart palpitations, and numbness of limbs after eating livestock and poultry products with such drug residues, which can be life-threatening in severe cases. Therefore, the United States and the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/54
CPCG01N30/02G01N30/06G01N30/54G01N2030/027G01N2030/062
Inventor 邵栋梁张洋朱晓慧周方
Owner ANHUI GUOKE TESTING TECH CO LTD
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