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

A technology for residues of various polypeptides and veterinary drugs, which is applied in the fields of food safety and analytical chemistry, can solve problems such as cumbersome processing methods, and achieve the effects of strong anti-interference ability, high recovery rate, and less dissolution of impurities

Inactive Publication Date: 2014-06-18
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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Example 1 Detection method of multiple polypeptide veterinary drug residues in pig liver

[0060] 1. Sample extraction: Weigh 2.0 g of the chopped pig liver sample into a 50 mL capped centrifuge tube, add an appropriate amount of anhydrous sodium sulfate, then add 10 mL of 1% trichloroacetic acid and 2 mL of acetonitrile, homogenize at 9 500 rpm After 2 min, centrifuge at 8500 rpm for 10 min at 4°C. The supernatant was transferred to another centrifuge tube through rapid quantitative filter paper. Add 15 mL of extract solution to the residue, ultrasonically extract for 10 min, shake for 10 min, and centrifuge at 8500 rpm for 10 min at 4°C. Combine the two extraction supernatants and wait for degreasing.

[0061] 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 lowe...

Embodiment 2

[0102] Example 2 Detection method of multiple polypeptide veterinary drug residues in milk

[0103] 1. Sample extraction: Weigh 2.0 g of milk sample into a 50 mL capped centrifuge tube, add an appropriate amount of anhydrous sodium sulfate and 1 mL of 6 mol / L hydrochloric acid, vortex for 1 min, then add 25 mL of extract solution, and ultrasonically extract for 10 min , extracted by shaking for 10 min, and then centrifuged at 8 500 rpm for 10 min at 4°C. The supernatant was filtered through rapid quantitative filter paper and then transferred to another centrifuge tube to be degreased.

[0104] 2, extract degreasing: with example 1.

[0105] 3. Purification of solid phase extraction column: same as example 1.

[0106] 4. Determination by liquid chromatography-mass spectrometry / mass spectrometer: same as example 1.

[0107] 5. Linear relationship test: Same as Example 1.

[0108] 6. Recovery rate and precision test: Accurately weigh milk samples that do not contain peptid...

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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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Patent Type & Authority Patents(China)
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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