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Method for detecting residual quantity of multiple polypeptidepolypeptide veterinary drugs in animal-derived food

A technology for a variety of polypeptides and veterinary drug residues, applied in the fields of food safety and analytical chemistry, it can solve problems such as cumbersome processing methods, and achieve the effects of strong anti-interference ability, high recovery rate and less impurity dissolution.

Inactive Publication Date: 2011-11-09
林维宣
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Most of the above methods for analyzing residues of polypeptide veterinary drugs are limited to the detection of single antibiotic residues, and the pretreatment methods are cumbersome

Method used

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  • Method for detecting residual quantity of multiple polypeptidepolypeptide veterinary drugs in animal-derived food
  • Method for detecting residual quantity of multiple polypeptidepolypeptide veterinary drugs in animal-derived food
  • Method for detecting residual quantity of multiple polypeptidepolypeptide veterinary drugs in animal-derived food

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Experimental program
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Effect test

Embodiment 1

[0059] 2. Degreasing the extract: Add 5-15 mL of n-hexane to the centrifuge tube of the extraction supernatant obtained in the previous step, vortex for 2 min, and centrifuge at 6 000 rpm for 5 min at 4°C. The upper layer (n-hexane layer) was removed, and the lower layer was purified by the solid phase extraction column.

[0060] 3. Solid-phase extraction column purification: Transfer the sample extract obtained above to the activated Oasis HLB purification column (activated with 6 mL of methanol and water), wash with 5 mL of water after loading the sample, and discard the effluent. Then elute with 5 mL of methanol, collect the eluate in a 10 mL graduated test tube, and control the flow rate during the entire solid phase extraction purification process to not exceed 2 mL / min. The eluent was blown dry with nitrogen on a water bath at 40°C, and the residue was dissolved in the standard sample solution to a volume of 1.0 mL, mixed evenly, and passed through a 0.22 μm microporous ...

Embodiment 2

[0102] 2, extract degreasing: with example 1.

[0103] 3. SPE column purification: same as example 1.

[0104] 4. Determination by liquid chromatography-mass spectrometry / mass spectrometer: same as example 1.

[0105] 5. Linear relationship test: Same as Example 1.

[0106] 6. Recovery rate and precision test: Accurately weigh milk samples that do not contain peptide veterinary drugs, add 3 different levels of peptide veterinary drug mixed standard solutions, the addition levels are shown in Table 5, and carry out 6 parallel samples for each concentration. Analysis was performed as described above in accordance with the above chromatographic and mass spectrometric conditions. The calculation results of recovery rate and coefficient of variation are shown in Table 7.

[0107] Table 7 Precision test results of peptide drugs added to milk (n=6)

[0108]

[0109] From the results in Table 7, it can be seen that the average recoveries of the three addition levels of polypept...

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Abstract

The invention relates to the fields of analytical chemistry and food safety, particular to a method for detecting the residual quantity of multiple polypeptide veterinary drugs in animal-derived foods. The method comprises the following steps of: processing a sample with a TCA(trichloroacetic acid) and acetonitrile system for depositing proteins; extracting with a carbinol and 0.1% formic acid aqueous solution system; purifying with a Oasis HLB solid phase extraction column; performing gradient elution with an Eclipse XDB-C18 analytical column in the presence of an acetonitrile and 0.1% formic acid aqueous solution used as a mobile phase; and then performing electrospray and positive ion scanning mode separation for finally detecting four polypeptides. The limits of the four polypeptides, namely colistin, bacitracin A, polymyxin B and virginiamycin M, are 25 micrograms / kilogram, 100 micrograms / kilogram, 250 micrograms / kilogram and 120 micrograms / kilogram respectively; the recovery rates of colistin, bacitracin A, polymyxin B and virginiamycin M are 74.9-88.1%, 76.2-89.0%, 76.6-81.2% and 77.3-86.9% respectively; and the coefficients of variation (CV%) of colistin, bacitracin A, polymyxin B and virginiamycin M are 5.7-15.1%, 7.2-15.7%, 6.0-8.0% and 9.5-18.6% respectively. The detection limit, the recovery rate, the accuracy and other technical indexes all meet related detection requirements at home and aboard.

Description

technical field [0001] The present invention relates to the field of analytical chemistry and food safety, in particular, it is a method for detecting residues of multiple polypeptide veterinary drugs in animal source food, especially bacitracin, polymyxin B, colistin, virginia A liquid chromatography-mass spectrometry / mass spectrometry method for the detection of four veterinary drug residues such as mycomycin. Background technique [0002] Polypeptide antibiotics ( peptides ) is extracted from the culture solution of Bacillus polymyxa or Bacillus aerogenes. Commonly used peptide antibiotics are bacitracin ( Bacitracin ), polymyxin B ( Polymyxin B ), colistin ( Colistin ), virginiamycin ( Virginiamycin )wait. Each peptide antibiotic is a compound with multiple components. Polypeptide antibiotics can respectively resist the infections of Gram-positive bacteria, Gram-negative bacteria, Pseudomonas aeruginosa, fungi, viruses, spirochetes, and protozoa, and can be us...

Claims

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Application Information

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IPC IPC(8): G01N30/72G01N30/06G01N30/08G01N30/34
Inventor 林维宣孙兴权蒋维旗董伟峰
Owner 林维宣
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